TECHNICAL REPORT
ON
STUDENT INDUSTRIAL WORK EXPERIENCE
SCHEME (SIWES)
UNDERTAKEN AT
ORILE AGEGE GENERAL HOSPITAL (O.A.G.H), 93, OLD
OTA ROAD, ORILE AGEGE, LAGOS STATE
BY
UTUBOR LUCKY OSAMUDIAME
MATRIC NO.: 13/50/0321
SUBMITTED TO
THE DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY
SCHOOL OF SCIENCE AND TECHNOLOGY
MOSHOOD ABIOLA POLYTECHNIC, PMB 2210,
ABEOKUTA, OGUN STATE,
IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF NATIONAL DIPLOMA IN SCIENCE LABORATORY TECHNOLOGY
NOVEMBER, 2014/FEBRUARY, 2015
CERTIFICATION
This is to certify that this report of the Student Industrial Working Experience Scheme (SIWES) programme was carried out by Utubor Lucky Osamudiame Student of Moshood Abiola Polytechnic, Abeokuta, Ogun State, Matric No.: 13/50/0321 at Orile Agege General Hospital (OAGH), 93, Old Ota road, Orile Agege, Lagos.
_____________________________ _____________________________
iStudent's Signature Supervisor's Signature
i
DEDICATION
This report is dedicated to the almighty God, the giver and sustainer of life, for His unconditional love and mercy granted to me throughout the period of my Industrial Training.
ii
ii
ACKNOWLEDGEMENTS
I give thanks to Almighty God, who gave me the gift of life, and made everything possible.
This acknowledgement would be incomplete if I fail to express my gratitude to my lovely family most especially my parents, Mr. and Mrs. Utubor, for their parental and financial support to my success. My appreciation also goes to Mrs. Egbuna Oluwafunmilayo, the HOD Clinical Laboratory Department, Orile Agege General Hospital and Mr. Odeyemi Abiodun Ayodele, my industrial based supervisor and other staff for all the imparts made to my Industrial Training.
iii
iii
REPORT OVERVIEW
The Student Industrial Work Experience Scheme established by the Federal Government of Nigeria was aimed at exposing student of higher institution to acquire industrial skill and practical experience in their approved course of study and also to prepare students for the industrial work situation which they are likely to meet after graduation. This technical report is based on the experiences gained during my four months of Industrial Training at Orile Agege General Hospital, Lagos State.
This report highlights how patients are being managed and also the several test carried out for patients such as: Full Blood Count (FBC), Packed Cell Volume(PCV),White Blood Cell Count, Differential Count, Stool Examination, Microfilaria, Widal (Typhoid test), Genotype, HIV, e.t.c. I was opportuned to work in five (5) sections which are Phlebotomy Section, Hematology/Immunohematology Section, Serology Section, Clinical Microbiology Section, and Chemical Pathology Section. These sections have exposed me to the precautions, rules and regulations of the laboratory, how to diagnose patients and how the tests are being analysed.
ivMost importantly, it describes the activities and my experience gained during the period of the training, it also stated the problems encountered and also gave suggestion for improvement of the scheme.
TABLE OF CONTENTS
iv
Title page
Certification …………………………………………………………................. i
Dedication ……………………………………………………………………… ii
Acknowledgements …………………………………………………………….. iii
Report Overview ………………………………………………………….......... iv
CHAPTER ONE
1.0 About the Industrial Training Fund (I.T.F)…………………………… 1-3
1.1 About SIWES (Student Industrial Training Fund)………………… 1
1.2 Scope……………………………………………………………….. 1-2
1.3 Aim and Objective of S.I.W.E.S…………………………………… 2-3
1.4 History and Background of Orile Agege General Hospital…………2-3
1.4 Organisation's Chart and Organogram……………………………..3
CHAPTER TWO
2.0 The Laboratory…………………………………………………………... 4-6
2.1 Introduction to the Laboratory…………………………………….... 4-5
2.2 Safety Rules in the Laboratory……………………………………… 5
2.3 Emergency in the Laboratory……………………………………….. 5
2.4 Hazardous Materials………………………………………………… 5
2.5 Hazardous Equipments…………………………………………...…. 5
2.6 Laboratory Equipments and their Uses……………………………… 5-6
CHAPTER THREE
3.0 The Laboratory Sections and Various Tests Performed……………….. 7-28
3.1 The Phlebotomy Department………………………………………… 7-11
3.2 Hematology and Immunohematolgy (Blood Bank) Section………… 12-16
3.3 Serology Section ………………………………………………….…. 17-19
3.4 Clinical Biochemistry Section …………………………………….… 19-22
3.5 Clinical Microbiology Section………………………………….……. 22-28
3.6 Microscopy, Culture and Sensitivity Tests …………………….…….. 28
CHAPTER FOUR
4.0 Summary, Challenges Encountered, Recommendation and
Conclusion………………………………………………………………..…. 29
4.1 Summary……………………………………………………….……... 29
4.2 Challenges Encountered ……………………………………………… 29
4.3 Conclusion ………………………………………………………….… 29
4.4 Recommendation …………………………………………………...… 29
CHAPTER ONE
1.0 ABOUT THE INDUSTRIAL TRAINING FUND (I.T.F)
In October 1971, the federal government established the Industrial Training Fund (I.T.F). In its policy statement no.1 published in 1973, a clause was inserted dealing with the issue of practical skills among the locally trained professionals in the tertiary institutions especially the Universities of Technology, Monotechnics, Polytechnics, Colleges of Education and Technical Colleges. Section 15 of the policy statement states clearly that "Great emphasis will be placed on assisting certain products of the post-secondary school system to adapt or orientate easily to their possible post graduation job environments", subsequently leading to the launch of a scheme know as the Student's Industrial work Experience Scheme(SIWES).
1.1 ABOUT SIWES
The S.I.W.E.S. was launched in 1973 by the Industrial Training Fund (I.T.F) as a programme designed to impart the undergraduate of the nation's tertiary institutions studying various professional courses with the practical methods of performing professional functions to real life situations on site, in the office or even the factory and how they apply themselves mentally, intellectually and physically in relation to what they have been taught in the classrooms theoretically. It works with the following professional bodies to function effectively across the tertiary institutions nationwide. These are the Nigeria University Commission (N.U.C), National Board for Technical Education (N.B.T.E.) and the National Commission for Colleges of Education (N.C.C.E.). Thus, equipping the students with the necessary skills and technical knowledge to make them highly competitive and professional individuals in the Labour Market
1.2 SCOPE
The scheme as conducted by the Industrial Training Fund (I.T.F) through their representative liaison units and offices situated within the various institution and in major cities or towns in Nigeria with the necessary industrial rudiments needed to corroborate, practicalize and then actualize the required technical knowledge. The Industrial Training experience not only puts them in real life situations buts also exposes their practical knowledge of the course of study, consequently perfecting this knowledge thereby producing very competent and versatile professionals.
1.3 AIM AND OBJECTIVE OF S.I.W.E.S
The aim of S.I.W.E.S is to bridge the gap between the level of knowledge acquired in tertiary institutions and the practical application of such knowledge in the field of work.
The Objectives are:
To provide an avenue for students in industries of higher learning to acquire industrial skills and experience in their course of study.
To prepare students for the work situations they are to meet after graduation.
To expose students to work methods and techniques in handling equipment and machinery that may not available in the educational institution.
To make transition from school to the world of work easier and enhance students contact for later job placements.
To improve student's interpersonal relationship with others in their field.
To prove students an opportunity to apply his/her knowledge in real work situation, thereby bridging the gap between college work and actual practice
1.4 HISTORY AND BACKGROUND OF ORILE AGEGE GENERAL HOSPITAL
Orile Agege General Hospital was established in 1999 by the Lagos State Government to enhance and promote the health services render by the Lagos State Health Service Commission (LSHSC).
It started as a health care centre and became a full operational General Hospital. Since establishment it has delivered quality services to the residents of Lagos State
COMPANY'S VISION
To be a world class human resource agency, ensuring the delivery of qualitative health care services for the people
COMPANY'S MISSION
To provide highly skilled and motivated staff with the right attitude to deliver efficient and effective health care to community
CORE VALUES
Medical excellence based on knowledge, skills and first rate human relations
Passions
Knowledge based hard work
Trust
Persistence
Imagination
Timeliness
Integrity
Professionalism
SCOPE OF SERVICE
Orile Agege General Hospital provides the following services to people:
Dental
Known evaluation, diagnoses, prevention and treatment of diseases, disorders and conditions of the soft and hard tissues of the jaw (mandible). The oral cavity, maxillofacial area and the adjacent and associated structures and their impact on the human body
ENT (Ear, Nose, and Throat)
The branch of medicine that specializes in diagnosis and treatment or ear, nose, throat and head and neck disorders.
Laboratory
This is where tests are done on clinical specimens in order to get information about the health of a patient as pertaining to the diagnosis, treatment and prevention of disease.
Obstetrics and gynecology
The entire scope of clinical pathology involving female reproductive organs and provision of care for both pregnant and non pregnant patients
Ophthalmology
This branch of medicine deals with the diseases and surgery of the visual pathways including the eyes, hair, and areas surrounding the eyes, such as lachrymal system and eyelids
Paediatrics
The branch of medicine that deals with the medical care of infants, children, and adolescents
Pharmacy
Charged with ensuring the safe and effective use of pharmaceutical drugs, the scope of pharmacy practice includes more traditional roles such as compounding and dispensing medications and it also includes more modern services related to health care including clinical services reviewing medications for safety and efficacy and providing drug information.
Radiology
The branch or specialty of medicine that deals with the study and application of imaging technology like X-ray and radiation to diagnosing and treating disease
Surgery
A medical specialty that uses operative manual and instrumental techniques on a patient to investigate and/or treat a pathological condition such as disease or injury to help improve bodily function or appearance or sometimes for some other reason.
VCT
Voluntary Counseling and Testing (VCT) for HIV
1.5 ORGANISATION'S CHART AND ORGANOGRAM
CHAPTER TWO
2.0 THE LABORATORY
2.1 INTRODUCTION TO THE LABORATORY
A laboratory is a facility that provides controlled conditions in which scientific researches, experiments, and measurements, may be performed. Hence the medical laboratory is a laboratory where tests are carried out on clinical specimens in other to get information about a patient's health.
There are three sections in the laboratory, they are; Clinical Microbiology section, Hematology/Serology section, and Clinical Biochemistry section. The overall significance of the laboratory diagnosis is that they guide towards the administration most effective therapy so as to restore a proper health on the patient. Laboratory safety precautions and ethics
2.2 SAFETY RULES IN THE LABORATORY
Every laboratory is expected to adopt a code of bio-safety principles and work practice which should be enforced and adhere to strictly by workers and visitors. All specimens coming into and from the laboratory are being assumed to be potentially infectious and harmful and that is why the below precautions are ensured to be taken to avoid contamination and laboratory hazard.
Avoid disrupting laboratory activities you must TURN OFF all cell phones and pagers: their use is prohibited.
All persons in laboratories, including students, staff, and visitors, shall wear safety glasses, goggles, or face shields at all times where potential eye hazards exist
Eating, drinking, chewing gum, and applying cosmetics are prohibited laboratory.
Do not store food or beverages in the same refrigerators or freezers with chemicals, biohazards, or radioactive materials.
Never conduct unauthorized experiments or engage in horseplay in a laboratory. Please immediately report any unsafe behaviour to the instructor.
Wear appropriate clothing. In particular, you must wear closed-toed shoes (i.e., NO sandals or flip-flops!) in the laboratory. If you have a long hair, tie it back. Avoid wearing dangling jewellery.
Wearing an iPod, Bluetooth, or any other device that interferes with hearing is not allowed.
Never pipette anything by mouth.
The work area must be kept clean and uncluttered. All chemicals should be labelled and stored properly.
The hazards of chemicals used should be known (e.g., corrosiveness, flammability, reactivity, stability, and toxicity).
Always pay attention to your surroundings and be aware of what others are doing. Always be courteous.
Remove contaminated gloves before touching common use devices (door knobs, faucets, equipment); discard gloves before leaving the laboratory.
Always wash hands and arms with antibacterial soap and water before leaving the laboratory.
In conclusion, maintaining safety in the laboratory largely rest on the shoulder of the laboratory workers. Adequate safety and good laboratory practice can be avoided irrespective of the location, staff strength and availability of sophisticated safety cabinets in the laboratory. What are required are highly standards of hygiene by the laboratory workers to achieve good results in their daily occupational practice.
2.3 EMERGENCY IN THE LABORATORY
Know where to find the nearest exit in case of fire or other emergency.
Know the whereabouts of the nearest fire extinguisher, fire blanket, first aid kit, eye wash equipment, shower and telephone.
In case of fire, clear out of the laboratory first, and then call an emergency number.
2.4 HAZARDOUS MATERIALS
Both liquid and dry chemicals can be flammable, poisonous, carcinogenic, etc. Pay attention to special instructions, such as to; work with a substance only in a fume hood.
Biological hazards include bacteria and body fluids, such as blood. Handle with appropriate care, and dispose of biological hazards as instructed.
Dispose of hazardous materials as instructed. Never put anything down the sink without checking with an instructor.
Clean up spills and broken glass. Don't handle broken glass with your bare hands. Use a broom and dustpan, and throw away all broken glass and disposable glass pipettes, coverslips, and other sharp or easily breakable glass in a container for glass disposal only. Notify the instructor immediately of all incidents.
2.5 HAZARDOUS EQUIPMENTS
If appropriate, turn off equipment that isn't being used.
Do not use a Bunsen burner unless instructed to do so.
Keep liquids and chemicals, especially flammable materials, well away from any heat source or electrical equipment.
If any electrical equipment is malfunctioning, making strange noises, sparking, smoking, or smells "funny," do not attempt to shut it off or unplug it. Get an instructor immediately. It is imperative that the instructor know of any equipment problems.
2.6 LABORATORY EQUIPMENTS AND THEIR USES
Microscope: Is used to examine samples and to analyze their contents that are not visible to the naked eye. It is used to count pathogen and other cells and to view under x10, x40, and x100 objectives.
Autoclave: For Sterilization
Centrifuge: Is used for spinning specimen e.g. urine to enable separation into constituents or components e.g. blood into serum and plasma.
Refrigerator: Provides suitable temperature for storage and preservation of reagents, unused media, blood samples etc.
Bunsen burner: Serves as the source of heat for sterilizing wire loop, surgical forceps and other metal instruments to be used for analysis.
Weighing Balance: Use for measurement.
Wire loop: It is used for streaking specimen on culture plates and it can also be used for making smear of samples on slides.
Lancet: It is a sterile needle used to prick the thumb for the collection of blood samples.
Capillary tube: It is used for the collection of blood samples to determine the packed cell volume.
Universal bottle: used for sample collection e.g. urine, stool, semen
Glass slide: It is used for the preparation of samples to be viewed directly under the microscope.
Sterile swab stick: Is used for the collection of samples to directly from the sight of infection e.g. Ear, nose, vagina, cervix, etc.
Sampling bottles: They are bottles used for the collection of blood samples e.g. universal bottle, fluoride oxalate bottle, Ethylene-Di-amine-Tetra acetic Acid bottle (EDTA), Lithium Heparin bottle, plain bottle.
Incubator: used for culturing or drying of microorganism.
Micro heamatocrit centrifuge machine: it is used to spin sample for the analysis of packed cell volume of blood sample.
Water bath: Use as heating apparatus
Micro haematocrit reader: used to read the packed cell volume in percentage.
Tourniquet: it is tightened on patient hand in the collection of blood sample in order to get a prominent vein before incision.
Needle and Syringe: It is used for the collection of blood samples.
Macro centrifuge machine: It is used for the separation of blood samples in order to get the plasma and also used for the separation of urine sample so as to get the supernatant and the specimen
Glucometer: used to check for the sugar level in the body with the aid of its strip.
Hematology analyzer: Is used for the analysis of Full Blood Count (FBC).
CHAPTER THREE
3.0 THE LABORATORY SECTIONS AND VARIOUS TESTS PERFORMED
3.1 THE PHLEBOTOMY DEPARTMENT
The Phlebotomy procedure facilitates obtaining good quality specimens on the correct patient for further analysis in the laboratory
3.1.1 IDENTIFICATION OF MATERIALS USED IN THE PHLEBOTOMY AND USES
Personal Protective Equipment
Phlebotomy Uniform: Serve as protective to the body
Disposable gloves/Hand Gel: For protection against spillage
Needles: Used to make incision
Needle holders: For holding needle or also known as venoject
Vacutainers, Vacutainer holder or Syringe: Serves as blood drawer
Sample bottles according to Order of Draw:
EDTA (Ethylene Diamine Tetra Acetic Acid)
Alkaline phosphatase - due to chelation of metallic cofactors which inhibits the enzyme activity. Potassium and sodium - due to the addition of potassium to the sample. Calcium and magnesium - these are chelated by the EDTA.
EDTA is also an unsuitable additive for samples requiring bacterial culture, since the chelation of the divalent cations inhibits the growth of bacteria. EDTA is sometimes used to prevent cells clumping in fluid samples requiring cell counts to accompany a cytology evaluation but it does not actually 'fix' the cells. A sample fixed in formalin or alcohol/ethanol is required for accurate cytology examination. EDTA is not suitable for samples requiring virus isolation in cell/tissue culture because it forms a gel when added to the cell culture medium and this disrupts the cell monolayer.
Plain (no anticoagulant)
Plain (or clotted) samples are used to provide serum for serology and most biochemistry or endocrine assays. Serum is plasma without fibrinogen since the fibrinogen has been used' in the formation of the clot.
Lithium Heparin
Lithium heparin accelerates the action of antithrombin III which neutralizes thrombin and thus prevents the formation of fibrin from fibrinogen (clot formation). This effect makes heparin samples unsuitable for determination of fibrinogen or clotting factors.
Lithium heparin is a standard anticoagulant used to obtain plasma for biochemistry analysis. Lithium heparin is the most suitable anticoagulant for the isolation of viruses in cell/tissue culture. This anticoagulant is not suitable for haematology as the heparin alters the cell morphology. Whilst measurement of haemoglobin and blood cell counts can be obtained using this anticoagulant an accurate white cell differential and morphology comments are not possible.
Fluoride/Oxalate
Fluoride/oxalate samples are used for glucose (and lactate) determination only. Sodium fluoride functions by stabilizing the blood cell membrane and inhibiting the enzyme systems involved in glycolysis, which prevents red blood cells metabolising any glucose present in the sample. For this reason it is the only suitable sample for accurate glucose analysis. Fluoride is a potent inhibitor of many enzymes and the inhibition of glycolysis tends to cause fluid shifts. Fluoride is a weak anticoagulant on its own, so potassium oxalate (another powerful enzyme inhibitor) is usually added to supplement its action. Other plasma or serum samples may be used for glucose analysis ONLY if the plasma/serum is separated from the cells within 1 hour of sample collection. Without an antiglycolytic agent, the blood glucose concentration decreases by approximately 0.56 mmol/l per hour at 25°C.
Sodium citrate
Sodium citrate is the standard anticoagulant for coagulation assays. Citrate functions by chelating calcium. The effect is easily reversed by the addition of calcium to the sample. Other anticoagulants are not suitable for coagulation assays as they interfere with coagulation factors. Citrate also inhibits ALP, ALT and interferes with the measurement of phosphate.
Tourniquet: Used to search vein. Prolonged venous occlusion can cause changes in concentrations of blood constituents. Therefore, the use of a tourniquet should be minimized. If a tourniquet is used to search for a vein, it should be released before withdrawal of blood begins. In any case, the use of a tourniquet should be limited to less than one minute.
70% Alcohol swabs: Used as a disinfectant for the area in which incision is to be made.
Micropore tape: To aid hemeostatis
Dental rolls
Adhesive dressing
Racks
Disposable waste bins
Cotton wool Pillow or other support: Used for supporting the arm for easy blood flow
3.1.2 STANDARD OPERATION PROCEDURE (S.O.P) FOR BLOOD COLLECTION
The frequent point of blood collection is usually from the vein (venipuncture). The materials for the patient's identity must be checked before attempting venepuncture. This must be carried out by asking the patient their Full Name and Date of Birth.
Check that this information corresponds with that on the Request form.
Any amendment to these details or any others on the Request form must be in accordance with the Directorate Policy.
Where patient details lack legibility, staff may write the correct details clearly next to those on the form without crossing out the original details.
If tests are requested that are unfamiliar and staff are unsure of the appropriate blood tubes for specimens check the list 'What tube guide' available at each workstation or, when necessary contact a qualified Biomedical Scientist in the appropriate department within the Pathology Laboratory.
Examine both arms of the patient and select the one that appears appropriate
Ensure that the patient is comfortable and that the arm is well supported and examine potential venepuncture.
Ask the patient to bare an arm, ensure that the arm is well supported and apply the tourniquet to the patient's arm, just above the elbow and tight enough to allow two fingers behind the strap.
Tighten the tourniquet a little more, taking care not to pinch the skin
Ask the patient to straighten their arm and clench their fist. This will make the vein more prominent.
If necessary rub the bend of the elbow to make the vein more visible.
Feel with a fingertip for the 'best' vein at the bend of the elbow rather than plunge the needle into a poor vein that looks 'alright'.
If this fails a suitable vein can often be found at the side of the arm on the elbow side.
It may be necessary, on occasion to take blood from the back of hand.
Apply the tourniquet above the elbow. The tourniquet is closed around the arm by inserting the plastic clip into the holder and then tightened appropriately by pulling the strap.
Ask the patient to straighten their arm and to make a fist in order to make the veins more prominent.
Feel with the fingertip if necessary to locate a suitable vein to puncture.
Ensure that equipment and blood tubes required are immediately within easy reach.
Remove the top plastic section of a Vacutainer multi-sample needle and screw thread into a Vacutainer needle holder.
Disinfect site with a 70% Isopropyl alcohol swab.
Leave for 30 seconds for the alcohol to evaporate and during this period assemble the blood tubes required.
Remove the cover from the multi-sample needle and discard into a clinical waste bin.
Keeping the needle holder and attached multi-sample needle in one hand use the thumb on the other hand to press on the vein just above the chosen entry point and pull the skin back slightly towards you to hold the vein firmly and stretch the skin over the chosen site.
With the needle holder and multi-sample needle almost parallel to the patient's arm and the needle bevel uppermost, gently push the needle into the chosen venepuncture site.
Once in the vein hold the needle-holder steady and gently push the cap of the appropriate blood sample tube onto the covered sample needle at the base of the inside of the needle-holder.
Blood should enter the sample tube and fill to the appropriate level indicated.
Remove the sample tube from the sample needle when full and attach another sample tube in required.
Blood samples must be gently mixed at the earliest opportunity to ensure anticoagulation effectiveness
As the last blood sample tube is filling slacken the tourniquet by pressing down on the release clip that is on the side away from the arm.
Withdraw the needle from the vein and quickly apply a clean pad of cotton wool.
Ask the patient to keep pressure on the cotton wool to stop further bleeding.
Discard the needle and holder into a Sharps bin.
Gently mix the sample tube(s).
If the patient is unable to maintain sufficient pressure on the vene-puncture site apply this pressure for them.
Remove the tourniquet from the patient's arm.
When bleeding from the venepuncture site has stopped apply Micropore tape tightly over the cotton wool.
The procedure is now complete and the patient can leave.
3.1.3 PHLEBOTOMY DIAGRAMS
3.2 HEMATOLOGY AND IMMUNOHEMATOLGY (BLOOD BANK) SECTION
In hematology section, the analysis is carried out using the whole blood sample of patient for diagnosis of hematological diseases and abnormalities. Blood samples are collected in EDTA bottle for analysis.
Immunohematology Section Also known as the blood bank performs tests to provide blood and blood products to patients for transfusion purposes. The blood bank technologist relies on the phlebotomist to perform identification of the patient without error, since patients will die if given the wrong blood type. The analyses carried out in these sections include: Packed cell volume, Full blood count, Erythrocyte sedimentation rate (ESR), Blood film for microfilaria and ABO/D (Rh) typing, Antigen typing, Blood grouping, Cross matching, respectively.
3.2.1 MATERIALS USED IN HAEMATOLOGY AND IMMUNOHEMATOLOGY SECTION
Pipette, Haematocrit centrifuge machine for PCV, Haematocrit centrifuge reader for PCV, Micro Haematocrit analyzer for Full Blood Count, Macro, Microscope, Microscopy slide, Electrophoresis machine, Cover slip, Bunsen burner, Plasticine, Sterile capillary tube, Wash bottle, Westergren tubes, Stop watch, Test tubes, Refrigerator, Racks, Various disposable waste bins, Tiles, Scissors, Ethylene diamine tetra acetic acid (EDTA), Leishman stain, Normal saline, Water, Antisera for blood group, Buffer solution, Oil immersion.
3.2.2 COMPLETE BLOOD COUNT (CBC) TEST
Introduction: The complete blood count of a blood sample helps to know the total cell in the whole blood. It determines the total haematocrit (HCT), hemoglobin (HGB), red blood cell (RBC) count, white blood cell (WBC) count, platelet count, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), differential (DIFF)-done on a blood smear
Aim: To deduce the total counts of all blood components
Equipments/Apparatus: Hematology analyzer, whole blood sample in an EDTA bottle.
Procedure: Blood sample was collected into an EDTA bottle (Lavender Stoppered Tube) through venipuncture and was mixed with anticoagulant by inverting the bottle gently 8 times. The blood sample was placed under the hematology analyzer sensitive probe. The probe button was pressed so that the probe can pick the sample into the machine for analysis. The result was displayed on the screen of the machine and then printed out.
Conclusion: The count of platelet, white blood cell and differentials, haemoglobin, granulocyte and all other cells in the blood samples was determined
3.2.3 PACKED CELL VOLUME (PCV) TEST
Introduction: The packed cell volume is the volume occupied by the packed red cell after a volume of anti-coagulated venous blood is fully centrifuged into plasma and red blood cell. The volume of packed cell is expressed as a percentage of the original volume of the blood.
Aim: To estimate the relative mass of red blood cells present in a blood sample in percentage volume.
Equipments/Materials: Haematocrit reader, Bunsen burner, Micro haematocrit centrifuge, Heparinised capillary tube, whole blood in an anticoagulant bottle (EDTA), Micro haematocrit reader, an absorbent cotton wool.
PLATE 2: MICRO HAEMATOCRIT READER
PLATE 2: MICRO HAEMATOCRIT READER
Procedures: The blood sample was collected into an EDTA bottle. The heparinized capillary tube was filled to 2/3 length of the tube from the blood sample and One end of the tube was sealed with flame using the Bunsen burner, then absorbent cotton wool was used in cleaning the tube before placing in the centrifuge. The sealed tube was placed in the micro- haematocrit centrifuge machine, thereby placing the sealed end outward to touch the base of the spinner. The sealed tube was spun in the haematocrit centrifuge at 12,000/13,000rpm for 5 minutes. The spun tube was placed on the micro haematocrit reader to read the result in percentage, positioned in slot so that the base line intersects the base of red cells and tube holder was moved so that the top line intersects the top of plasma, then knob was adjusted so that the middle line intersects the top of red cell.
The percentage packed cell volume on the scale was read.
Result: Adult: Normal range for male 37-50%
Normal range for female 35-45%
Children: Normal Range 29-41%
Conclusion:
Factors affecting the accuracy of PCV are; Unsteady power supply, Poor blood sample collection, Parallax error while reading the result on the haematocrit reader, Incorrect blood to anticoagulant ratio, Over spinning of the blood in the centrifuge, Lysing of the blood by flame or delay in spinning
Bio- medical significance: Low PCV value indicate shortage of blood
3.2.4 ERYTHROCYTE SEDIMENTATION RATE (ESR)
Introduction: Erythrocyte Sedimentation rate test is used to detect inflammation infections, cancer, and autoimmune diseases; it is also use to monitor whether an illness is responding to treatment. It is a screening test so cannot be used to diagnose a specific disorder
Aim: To determine the rate of inflammation of blood using Westergren method.
Equipment/Apparatus: Sodium Citrate bottle, ESR rack, Westergren glass rod.
Procedure: Blood sample was collected into an EDTA bottle and inverted 8 times to enable the action of the additive and then poured into the Westergren tube containing sodium citrate and was mixed gently again by inverting the tube 8 time. The Westergren glass rod was forced into the tube and blood rises to zero mark. The sample was placed in a vertical stand on the ESR rack and timed for one-hour. The result was recorded just after one-hour in millimeters
Result: The normal range of the sedimentation in adult
Men under 50 years………………………..less than 15mm/hr
Men over 50 years………………………….less than 20mm/hr
Women under 50 years………………..........less than 20mm/hr
Women over 50 years……………………….less than 30mm/hr
The normal range of the sedimentation in children
New born ………………………………………… less than 2mm/hr
Neonatal to puberty ……………………………..3- 13mm/hr
Conclusion: The ESR test does not diagnose a particular kind of infection but rather tell if further medication should be given. It is usually done to indicate if there are abnormal proteins in the body
3.2.5 BLOOD GROUPING AND GENOTYPING TEST
Introduction: Blood grouping of the A B O system is determined with Anti-A, Anti-B, and Anti-D sera, which form agglutination complex with antibodies found in the blood sample.
Aim: To determine the group and the rhesus of a patient's blood
Equipments/Materials: Clean free grease tile, Pasteur pipette, Whole blood sample in an EDTA bottle, distilled water, applicator stick, test tube rack, electrophoresis machine and tank, clean white tile, cotton wool, applicator stick, cellulose filter paper, gloves.
Reagents: Anti-A, Anti-B, Anti-D sera, Buffer for balancing, normal saline
Procedure:
For blood Grouping: The blood sample was collected into an EDTA bottle through venipuncture. 10ul of blood was placed 3 spots on the tile with the aid of Pasteur pipette. The antisera A, B and D were placed carefully on each spots, ABO of the grouping system on the tile respectively and an applicator stick was used to thoroughly mix the drop of blood with the anti-sera one after the other without contamination. The tile was gently rocked from side to side for 3 minutes to allow agglutination occurrence, then result was observed.
For Genotyping: Cells were washed two to three times in a test tube containing normal saline after which, a drop of washed cells were placed on a tile. This is followed by the hemolysis of blood on the tile and the placement of AS and AA control using applicator stick, after making sure that the buffer inside the electrophoresis tank covered the electrode, the cellulose paper was placed on the tank, which is then covered and mains (current) switched on. Reading was recorded after 5-10mins
Result: The result for blood genotype was taken by studying the movement and separation of hemoglobin molecule.
Conclusion: The result was observed according to the agglutination that occurred in each spots on the tile. Anti D determines the present of the rhesus 'D' factor in blood group.
Factors that affect blood grouping are; wrong labeling of spot and confusion of anti-sera with spots, Contamination of test card or tiles with detergents, Expired anti-sera
Bio-medical significance; Blood transfusion, Blood compatibility, Antenatal screening#
3.3 SEROLOGY SECTION
Tests done in this department are designed to detect the body's response to the presence of bacterial, viral, fungal, parasitic and other conditions which stimulate detectable antigen-antibody reactions in a test system to aid in the diagnosis of the patient. Most tests performed in this section are carried out under the principles of Immunoassay, some of them are; Cold agglutinins (CAG) - specimen must be kept warm, Rheumatoid arthritis (RA), VDRL, to diagnose syphilis, Pregnancy Testing, Widal
3.3.1 HBs.Ag TEST FOR HEPATITIS, VDRL (VENERAL DISEASES RESEARCH LABORATORY) TEST FOR SYPHILIS USING STRIPS
Introduction: HBsAG is a rapid immunochromatographic test for the qualitative detection of Hepatitis B surface Antigen in human serum/plasma, it can be used for prenatal or transfusion screening, and during acute infection or chronic carriage of the Hepatitis B virus. Early detection of infection is essential for rapid initiation of adequate treatment.
VDRL test is a screening test for syphilis. It measures substances called antibodies that body may produce if it comes in contact with the causative agent of syphilis, which is called Treponema pallidium
Aim: To determine the presence or absence of hepatitis and syphilis in the body system.
Materials: HBsAG Test strips, VDRL test strip EDTA bottle, Centrifuge, clean test tube
Specimen: Serum.
Procedure: The patient blood sample was collected into a plain bottle through venipuncture. The blood sample was spun in a centrifuge for 5 minutes, after spinning the serum was separated carefully into a clean test tube by the use of Pasteur pipette and then test strip was immersed vertically into the serum for 10 minutes. The observation was taken after 10mins.
Result: Appearance of a line at the Control region and another at the Test indicates positive result, while an appearance of a line at the Control region only, indicates negative result. When there is no appearance of any line, means the test in invalid and as to be redone using new kits
3.3.2 BLOOD PREGNANCY AND URINE PREGNANCY TEST, USING TEST STRIP.
Introduction: A pregnancy test is done to determine if a woman is pregnant, pregnancy hormone called the Human Chorionic Gonadotrophin (HCG) in to the blood and urine. Pregnancy test detects the hormone HCG and confirms the pregnancy.
Aim: to determine the presence of pregnancy hormone (HCG) in the blood and urine.
Materials: pregnancy test strip, plain bottle, needle and syringe, wet swab, cotton wool, centrifuge, clean test tube.
Specimen: blood (serum) and urine
Procedure for Blood Pregnancy Test: Patient's blood was collected through venipuncture into plain bottle, blood sample was spun in a centrifuge for 5 minutes, and the serum was separated carefully into a clean test tube by the use of Pasteur pipette. The pregnancy test strip was immersed vertically into the serum for 5 minutes. The strip was removed and the reaction was observed.
Procedure for Urine Pregnancy Test: The patient's urine sample was collected into universal sterile bottle and the pregnancy test strip saw immersed into the urine for 3 seconds, then removed and left for 5mins and the result was observed.
Result: An appearance of a line at the Control region and another at the Test indicates positive result, while an appearance of a line at the Control region only, indicates negative result. When there is no appearance of any line, means the test in invalid and as to be redone using new kits
3.3.3 WIDAL TEST
Introduction: Typhoid fever is an infectious disease caused by the Salmonella typhi, it is diagnosed by Widal test which employs an antigen-antibody reaction to screen for the presence of Salmonella typhi and paratyphi antibodies in the sample serum.. the organism is transmitted by water or food contaminated by faeces of typhoid victims or carriers, that is a person who harbor it without showing signs and symptoms.
Aim: To investigate the presence of Salmonella typhi and paratyphi in the serum of patient
Materials: Test card/white tiles, Pasteur pipette, centrifuge, antigen kit and stop watch
Procedure: 3-5ml of blood was collected from the patient through venipuncture into a plain bottle and the blood was spun at 3000rev per min for 5minutes so as to separate out the plasma. A dropper was used to carefully drawn the antigen kits and a drop was placed on each of the test card in pairs of four spots labeled O, OA, OB, OC and H, HA, HB, HC and a drop of serum was carefully added into the antigen respectively with the aid of Pasteur pipette and mixed together with the aid of an applicator stick the test card was rocked gently, the rate of reaction and agglutination was observed at an interval of 30sec, 1min, 2mins, and 5mins
Result
Reactive: visible agglutination on spot H and others indicate the present of Salmonella antibodies
Non reactive: no visible agglutination indicates absence of Salmonella antibodies
Widal test: Positive
Highly reactive……………………………….1:320 (agglutination reaction before 60 seconds)
Very reactive……………………………….1:160(agglutination reaction before 120 seconds)
Reactive ………………………………………1:80(agglutination reaction before 180 seconds)
Widal test: Negative
Non significant…………………………………1:40
Non significant………………………………….1:20
Not reactive…………………………………….Nil
3.3.4 RETROVIRAL TEST
Introduction: This is the diagnosis for Human Immunodeficiency Virus, an infectious agent that causes Acquired Immunodeficiency Syndrome (AIDS), a disease that leaves a person vulnerable to life threatening infection. HIV transmission occurs when a person id=s exposed to body fluids infected with virus, such as blood, semen, vaginal secretions and breast milk.
Aim: To investigate the presence of HIV 1 and 2 in patient's blood
Materials: Determine kits, Unigold kit, Stat pack Buffer, Plain bottle, pipette
Specimen used: Serum
Procedures: The blood sample, collected in a plain bottle was centrifuged at 3000rev/min to allow separation. The serum was picked with a pipette and two drops was placed on the sample pad of the determine kit and allowed to penetrate then left for 15min. If result proves positive, the Unigold kits would be used following the same procedure. After using Unigold to confirm the result and proves negative the Stat pac kit would be used to as a confirmer.
Result: The appearance of a line at the Control region and another at the Test region indicates a Positive result, while the appearance of a line on the Control region only, and indicates a Negative result.
3.4 CLINICAL BIOCHEMISTRY SECTION
This section deals with chemical analysis of serum or plasma in which many diseases of the major organs systems can be diagnosed such as heart attacks, hepatitis, renal failure, diabetes, Liver function, etc Blood sample samples may collected into the Serum Separator Tube or Lithium Heparin. Test performed in this department are:
Blood Glucose; FBS, FBS2HPP, RBS, OGTT
Blood lipids (fat) Cholesterol level.
Electrolytes - sodium, potassium, CO2- (Bicarbonate), and chloride
Uric acid
Creatinine and Blood Urea Nitrogen (BUN)
Liver function tests -AST, ALT, alkaline phosphatase, and bilirubin.
3.4.1 MATERIALS AND EQUIPMENTS USED IN CLINICAL BIOCHEMISTRY SECTION
Personal Protective Equipments (PPE), Blood collection materials Different tubes like; Lithium Heparin, Serum Separator and Fluoride Oxalate tubes, Chemical reagents and detergents, automated machines, centrifuge, Glucometer and Accu check.
3.4.2 BLOOD GLUCOSE
Test Overview: A blood glucose test measures the amount of a type of sugar, called glucose, in your blood. Glucose comes from carbohydrate foods. It is the main source of energy used by the body. Insulin is a hormone that helps your body cells uses the glucose. Insulin is produced in the pancreas and released into the blood when the amount of glucose in the blood rises.
Normally, your blood glucose levels increase slightly after you eat. This increase causes your pancreas to release insulin so that your blood glucose levels do not get too high. Blood glucose levels that remain high over time can damage your eyes, kidneys, nerves, and blood vessels.
There are several different types of blood glucose tests.
Fasting blood sugar (FBS) measures blood glucose after you have not eaten for at least 8 hours and at most 14 hours. It is often the first test done to check for prediabetes and diabetes Materials/Reagents: Fluoride Oxalate and other blood collection equipments, Centrifuge, Insulin kit (NORUDIA® Insulin) Liquid reagent, automated machine, Glucometer or Accu Check.
Procedure: Patients is ensured to have fasted for the required period of time before sample collection into a Fluoride Oxalate tube, and then the blood samples are centrifuged in order to obtain plasma. The plasma obtained was decanted into a small cap which is then transported into the machine for analysis. The results are then deduced which is measured in mg/dl
Normal Result: Normal result is less than or equal to 100mg/dl
2-hours post-prandial blood sugar measures blood glucose exactly 2 hours after you start eating a meal. This is not a test used to diagnose diabetes.
Procedure: After performing the same procedure for FBS sample collection, patients are then asked to return 2hrs as soon as they start eating, then another venepucture is made and the same procedure is repeated.
Result: Normal results is less than 140mg/dl for people age 50 and younger; less than 150mg/dl for age 50-60; less than 160mg/dl for age 60 and older.
Random blood sugar (RBS) also known as casual blood glucose test. RBS measures
Blood glucose regardless of when you last ate. Several random measurements may be taken throughout the day. Random testing is useful because glucose levels in healthy people do not vary widely throughout the day. Blood glucose levels that vary widely may mean a problem.
Materials/Reagents: Same materials as FBS.
Procedure: Glucometer or Accu Check are mostly used to perform this test. The Glucometer corresponding code strip is inserted and loaded, Patient's thumb is disinfected using a70% alcohol pad and pricked with lancet. The blood is wiped off in order to avoid sampling error, pressure is applied below to enable blood flow again and the blood is placed on the strip. The result is then taken
Normal Results: 80-120mg/dl before meals and 100/140mg/dl after meals
Oral glucose tolerance test is used to diagnose prediabetes and diabetes. This test is a series of blood glucose measurements taken after you drink a sweet liquid that contains glucose. This test is commonly used to diagnose diabetes that occurs during pregnancy (gestational diabetes). This test is not commonly used to diagnose diabetes in a person who is not pregnant.
Materials/Reagents: : Fluoride Oxalate and other blood collection equipments, Centrifuge, Insulin kit (NORUDIA® Insulin) Liquid reagent, automated machine, glucose solution
Procedure: Patient is asked to take the required gram of sugar solution before sample collection. Samples are collected five times at 30min intervals and lastly collected once 1hr after the last sample collection of 30min intervals. The result is the deduced by the machines.
Results: 75g of glucose; fasting 92mg/dl or more; 1hr 180mg/dl or more; 2hrs 153mg/dl or more
100g of glucose; more than or equal to 140mg/dl
Haemoglobin A1c (also called glycated haemoglobin) measures how much sugar (glucose) is stuck to red blood cells. This test can be used to diagnose diabetes. It also shows how well your diabetes has been controlled in the last 2 to 3 months and whether your diabetes medicine needs to be changed.
Materials/Reagents: : Fluoride Oxalate and other blood collection equipments, Centrifuge, Glycohemoglobin kit (NORUDIA® HbA1c) , automated machine, glucose solution
Procedure: Same procedure as FBS
Result: The result of your A1c test can be used to estimate your average blood sugar level. This is called your estimated average glucose, or eAG which ranges: (126, 154, 183, 212, 240, and 289) mg/dl
ELECTROLYTES TEST
Test Overview: Electrolytes are minerals found in the body tissues and blood in form of dissolved salts which helps transfer nutrients into body cells and waste of them. Electrolytes also maintain a healthy water balance and help stabilize body's acid/base (pH) level. The main electrolytes in the body are; Sodium, and Potassium others are; CO2- (Bicarbonate), and chloride
3.4.4 URIC ACID TEST
Test Overview: This is a kidney or liver function test, which measures the amount of uric acid present in a blood sample. It is produced from the natural break down of body's cells and from food eaten and then filtered out by the kidneys and passes out of the body in urine but if too much is being produced in the body or the kidney is unable to filter them normally, it becomes high and may cause solid crystals from within joint, which may lead to a painful condition called gout. These uric crystals can build up in joint and nearby tissues, thereby forming hard lumpy deposits called tophi
Results: Normal Range; in men 3.4 – 7.0mg/dl, in women 2.4 – 6.0mg/dl, in children 2.0 – 5.5 mg/dl
3.4.5 BLOOD UREA NITROGEN AND CREATININE TEST
Test Overview: This test is used to depict the function of kidney. Blood Urea Nitrogen and Creatinine test can be used together to find the BUN-to-Creatinine ratio and when these substances are high in the blood it may lead to heart failure, dehydration. Blood urea nitrogen (BUN) measures the amount of Nitrogen in your blood that comes from the waste product urea. Urea is made when protein is broken down in your body. Urea is made in the liver and passed out of your body in urine.
Normal Results: Blood Urea Nitrogen (BUN); Adults: 10-20mg/dl, Children: 5-8mg/dl
Blood Creatinine; Men: 0.6-1.2mg/dl, Women: 0.5-1.1mg/dl, Children; 0.4 to 1.0mg/dl
3.4.6 LIVER FUNCTION TEST
This is a group of blood tests that detects inflammation and damage to the liver and also check how well the liver works. Liver function test includes ALT (Alanine amino trans ferase) , AST (ASpartate aminotransferase others are PT, INR, albumin, bilirubin ALP (Akaline phosphatase)
3.5 MICROBIOLOGY SECTION
Microbiology involves the study of microbes. Although, microorganisms are generally beneficial and essential to life some are, however, pathogenic and cause infectious diseases. The diagnostic microbiology laboratory is engaged in the identification of infectious agents. These infectious agents are broadly classified as viruses, bacteria, mycostic agents and parasites (Protozoans and Helminthes) also this section analyses body fluids and tissues for the presence of pathogenic microorganisms primarily by means of culture and sensitivity (C&S). Results of the C&S tell the physician the type of organisms present as well as the particular antibiotic that would be most effective for treatment
3.5.1 BASIC RULES FOR WORKING IN THE MICROBIOLOGY LABORATORY,
While working in the laboratory, it is important that you must adhere to the following basic rules;
Be methodical and orderly in habits; keep the work area clean especially before leaving the laboratory and disinfectant it thoroughly at the end of day.
Wash hands frequently with soap and water
Before leaving the laboratory, place the discarded glassware into the designated place.
Cultures are kept under incubation and should be inspected in the morning and findings must be carefully.
Send the laboratory reports promptly. In case of emergency a special report is dispatched or communicated by telephone.
3.5.2 GENERAL REMARKS
All specimens for culture must be collected prior to the therapy. If the patient is on antibiotic, inform the microbiologist so that he/she can take measures.
Collect the specimen in adequate amount from the infectious site. This usually instructed by the physician.
Always use the sterile bottle to transporting the specimen.
All specimen must be accompanied by a request slip with complete information, h on the patient name, age, sex, hospital number, source of specimen and clinical information is very important in order to choose the appropriate medium for the culture.
Enter the details in the laboratory register and performed direct examination of the specimen before choosing the media for the culture.
All containers used for holding microbiological specimens must be sterilized before used. Such as test tube, culture tube with and without cap, and plates, container to collect sputum specimen, blood specimen for microbial culture, penicillin bottles for collection of spinal fluid and other specimen container (universal bottle) for collection of urine specimen and stool specimen.
3.5.2 MATERIALS/APPARATUS USED IN MICROBIOLOGY SECTION
Inoculating loop, Bunsen burner, Incubator, Weighing balance, Spatula, Microscopy slide, Cover slip, Staining rack, Medium plates, Sensitivity disc, Forcep, Cotton wool swab.
3.5.3 INTRODUCTION LABORATORY GROWTH MEDIA
These are classified into six types: (1) Basal media, (2) Enriched media, (3) Selective media, (4) Indicator media, (5) Transport media, and (6) Storage media.
1. Basal Media. Basal media are those that may be used for growth (culture) of bacteria that do not need enrichment of the media. Examples: Nutrient broth, nutrient agar and peptone water. Staphylococcus and Enterobacteriaceae grow in these media.
2. Enriched Media. The media are enriched usually by adding blood, serum or egg. Examples: Enriched media are blood agar and Lowenstein-Jensen media. Streptococci grow in blood agar media.
3. Selective Media. These media favour the growth of a particular bacterium by inhibiting the growth of undesired bacteria and allowing growth of desirable bacteria. Examples: MacConkey agar, Lowenstein-Jensen media, tellurite media (Tellurite inhibits the growth of most of the throat organisms except diphtheria bacilli). Antibiotic may be added to a medium for inhibition.
4. Indicator (Differential) Media. An indicator is included in the medium. A particular organism causes change in the indicator, e.g. blood, neutral red, tellurite. Examples: Blood agar and MacConkey agar are indicator media.
5. Transport Media. These media are used when specie-men cannot be cultured soon after collection. Examples: Cary-Blair medium, Amies medium, Stuart medium.
6. Storage Media. Media used for storing the bacteria for a long period of time. Examples: Egg saline medium, chalk cooked meat broth. The survival and growth of microorganisms depend on available and a favorable condition environment. Culture media are nutrient solutions used in laboratories to grow microorganisms. The method for the preparation of basic microbiology media is given below. Sterilization is at 121c for 15minutes.pH values are 7.0 unless stated otherwise.
3.5.4 COMMON MEDIA IN ROUTINE USE
CLED Agar (Cysteine Lactose Electrolyte Deficient): is a non-selective differential plating medium for the growth and enumeration of urinary tract microorganism.
Preparation: 36.0g of medium is suspended in one liter of distilled water, slowly heated and frequently stirred. Boiled for a minute and sterilized at 121OC for 15minutes and poured into Petri dishes. Plates were inverted when solidified for storage purposes and to avoid moisture
MacConkey Agar. Most commonly used for enterobac teriaceae. It contains agar, peptone, sodium chloride, bile salt, lactose and neutral red. It is a selective and indicator medium:
Selective: as bile salt does not inhibit the growth of enterobactericeae but inhibits growth of many other bacteria.
Indicator: medium as the colonies of bacteria that ferment lactose take a pink colour due to production of acid. Acid turns the indicator neutral red to pink. These bacteria are called 'lactose fermenter', e.g. Escherichia coll. Colourless colony indicates that lactose is not fermented, i.e. the bacterium is non-lactose fermenter, e.g. Salmonella. Shigella, Vibrio.
Preparation: 50g of the Agar was suspended and measured into one liter of purified water and mixed thoroughly and was heated with frequent agitation, then boiled for one minute to completely dissolve the powder. The Agar was autoclaved at 121OC for 15minutes.
Chocolate Agar or Heated Blood agar. Prepared by heating blood agar. It is used for culture of pneumococcus, gonococcus, meningococcus and Haemophilus. Heating the blood inactivates inhibitor of growths.
Preparation: 2litres of distilled water was added to 144g of agar powder. Autoclaving was done at 121OC for 15minutes and cooled till 45OC then 5% of defribrinated blood was added. Heated slowly and evenly to 65OC, cooled till 45OC and poured into plates
Blood Agar. Most commonly used medium. 5-10% defibrinated sheep blood is added to melted agar at 45-50°C. Blood acts as an enrichment material and also as an indicator. Certain bacteria when grown in blood agar produce haemolysis around their colonies. Certain bacteria produce no haemolysis. Types of changes:
(a) Beta (β) haemolysis. The colony is surrounded by a clear zone of complete haemolysis, e.g. Streptococcus pyogenes is a beta haemolytic Streptococci.
(b) Alpha (α) haemolysis. The colony is surrounded by a zone of greenish discolouration due to formation of biliverdin, e.g. Viridans streptococci, and
(c) Gamma (γ) haemolysis, or, No haemolysis. There is no change in the medium surrounding the colony,
Nutrient Broth. 500 g meat, e.g. ox heart is minced and mixed with 1 litre water. 10g peptone and 5 g sodium chloride are added, pH is adjusted to 7.3. Uses: (1) As a basal media for the preparation of other media, (2) To study soluble products of bacteria.
3.5.5 DISPOSAL
Once the Petri dishes have been taped shut, they should not be opened again. All microorganisms grown during the experiment should be killed before discarding. The best way to dispose of bacterial cultures is to pressure sterilize them in a heat stable biohazard bag. If autoclaves or pressure cookers are not available or large enough to make this convenient, an alternative is to bleach the plates. Saturate the plates with a 20% or "1 in 5" household bleach solution (in other words, 1part bleach and 4 parts water). Let them sit and soak overnight in the bleach solution before disposing of them. Please note that the bleach solution is corrosive and needs to be thoroughly removed afterwards. In addition, the plates can be incinerated if access to an incinerator is available.
3.5.6 PRECAUTIONS TAKEN WHEN PREPARING MEDIUM
Do not talk when pouring medium on plates and when culturing the sample on plates to avoid contaminants as a result of unwanted bacterial or enzymes through saliva.
The degree at which we incubate any cultured sample is always at 37c to avoid the death of the microorganism.
3.5.7 GRAM STAINING
Introduction: In this section, the staining of bacteria as a means for identification is done. Bacteria are being identified as Gram-Negative or Positive on the basis of their cell wall thickness after staining. Gram positive bacteria hold the dye and appear purple while Gram-Negative bacteria release the dye and appear red.
Aim: To identify the gram-negative and the gram positive bacteria.
Apparatus: Stop watch, Normal saline, Clean grease free microscopic slide, Gentian violet, Lugol's iodine, 95% ethyl alcohol, Neutral red, Microscope, Sterilized inoculating loop.
Procedure: The organism was isolated and smeared using the sterilized inoculating loop in a drop of normal saline on a clean grease free microscopic slide. It was left to Air-dry.
The smear was placed on a staining rack, and was flooded with Gentian violet solution for 30seconds. It was rinsed with water. The smear was again flooded with Lugol's iodine for 30 seconds. It was rinsed with water and the smear was decolorized with 95% ethyl alcohol for 30 seconds, It was rinsed with water again.
The decolorized smear was counter-stained by flooding with neutral red for 30 second and was rinsed with water. The back of the slide was cleaned with cotton wool and allowed to Air-dry. The slide was mounted on the microscope after air-drying and was examined under ×100 immersion objectives. The result was recorded.
Sensitivity Test (Result): If the bacteria are gram positive, a positive sensitive disc is used while a negative sensitive disc is used for gram negative bacteria. A pure colony was sub-cultured on a Nutrient medium and sensitive disc was picked with the aid of a sterile forceps, and placed on the medium, then the plate was incubated for 24 hours at 37OC. The plate was read after 24hours of incubation to observe zone of inhibition and resistance. Sensitivity test is also done for pathogens that grow on the media by taking a colony of the organism, streak on the sensitivity agar and add antibiotics discs and incubate for 24hours at 37OC.
3.5.8 PROCEDURE FOR AUTOCLAVING
The inner element was filled with water and allowed to cover the surface of the element, The medium were arranged in the sample bucket and the machine was covered and screwed tightly by holding the screw opposite each other and the wing nut was screwed tightly.
The switch on button was on and also the outlet valve was opened down so as to increase the temperature of the steam on the workload.
It was sterilized at 121 C for 15minutes. The safety valve was closed at 120 C and at 121 C, button was switched off. The medium were allowed to cool for 47 C before pouring in the petridish
Principle behind Autoclaving.
The principle behind these sterilization methods is based on the temperature and the type of autoclaving operation performed. Autoclaving operation at 121 C is referred to as culture media autoclave.
3.6 MICROSCOPY, CULTURE AND SENSITIVITY TESTS
Microscopy involves the examination of specimen under the microscope, Culture refers to the microorganisms that grow on the culture medium after inoculation and incubation while sensitivity tests determines the antibiotics which will be administered to the patients.
The cultured plates are incubated for 24hours at 37oC to facilitate the growth of the organism and chocolate plates were incubated in a candle jar to facilitate the growth of both aerobic and anaerobic microbes while other plates were incubated aerobically.
3.6.1 STOOL ANALYSIS
Introduction: Stool analysis involves the collection and analysis of faecal matter to diagnose the presence or absence of a medical condition. During the outbreak of cholera or diarrhea,food microbiologist collect stool sample into sterile universal bottle from victims for faecal examination in the laboratory.
Aim: To determine cysts and ova in the stool of a patient.
Materials/Apparatus: Clean microscopic slide, drop of saline, binocularmicroscope, swabstick, coverslip, and a loop.
Procedures: Sample was collected into a bottle, a drop of normal saline was mounted on a clean microscopic slide and mixed it with a small portion of fresh faeces with a loop. The slide was covered with a cover slip and the viewed under the low-power objective by using *10 and *40 for clear viewing.
Results: The results were viewed in two ways which are microscopic and macroscopic;
Macroscopic-----------------Hard, dark ,brown stool formed with no mucus or blood seen.
Microscopic-----------------Present of Ascaris lumbricoides .
3.6.2 URINE MICROSCOPY CULTURE AND SENSITIVITY (M/C/S)
Urine for microscopy culture and sensitivity is an array of tests performed on urine samples to examine the presence or absence of cells such as; epithelial cells, pus cells, red blood cells, yeast cells, crystals, and bacteria. It is one of the most common methods of medical diagnosis.
Aim: To identify parasites and Bacteria cell in the urine of individual.
Materials: Microscope, microscopy slide, centrifuge, inoculating loop, urine sample, medium plates; Macconkey agar and CLED agar, Nutrient agar, Gram staining reagents, Sensitivity disc, Incubator, source of heat.
Procedures:
Microscopy: 5ml of the urine was transferred for spinning in a bench centrifuge at 30000rev/min for 5minutes. The sediment was concentrated in the test tube by decanting off the supernatant.
A small drop of the sediment was placed on a clean slide covered with a cover slip as wet preparation and then mounted on a microscope. The slide was examined using x40 objective lens.
Culture: A loopful of the sample was used to make a streak on the Agar plates.
The plates were then incubated for 24hours at 37OC. The culture media were removed from the incubator after 24hours and visible bacteria growth was read and recorded.
Gram Staining: Primary and secondary gram staining was done on the smear of a colony of the bacteria. This is to identify if the bacteria is Gram positive or Gram negative.
Results:
Macroscopy: The following parameters are examined
Appearance: Clear, turbid ,slightly turbid,
Colour: Yellow, straw, Amber yellow, pale yellow.
Microscopy: Presence or absence of ; Pus cells, Epithelia cells, Red blood cells, Yeast cells, Bacteria cells and Crystals can be seen in the wet preparation.
Culture: The following Bacteria can be isolated from Urine samples; Klebsiella spp, Staphylococcus aureus, Coniforms
3.6.3 EYE, EAR NOSE AND THROAT, WOUND AND PUS SWAB
Procedure: These samples are collected using a dry sterile cotton wool swab; they are then inoculated into Chocolate and Macconkey agar. Gram staining is then carried out on each specimen. Pathogens that are likely to be observed include;
Wound and Pus swab: Staphylococcus aureus, Sreptococcus pyogenes, Clostridium perfrigens
Eye swab: Haemophilus influenza, Pseudomonas aeruginosa, Betahaemolytic streptococcus etc.
Ear swab: Escherichiacoli, Proteussp etc.
Throat Swab: Throat cultures are submitted primarily for the detection of Group A Streptococcus. When obtaining the specimen, depress the tongue with a tongue blade, and swab the tonsillar pillars and behind the uvula including any inflamed or purulent sitesAvoid touching the tongue, cheeks or teeth. Immediately place the swab back into the culturette sleeve and crush the ampule
3.6.4 HIGH VAGINAL, URETHRAL AND ENDOCERVICAL SWAB.
Test Overview: This is use to detect the causative organisms of female reproductive system infections and their sensitivity to antibiotics.
Procedure: A swab stick was used to collect specimen from the affected area and then inoculated into sterile media which include Chocolate and MacConkey agar. These plates were incubated at 37c for 24hours and were examined for any pathogenic growth, if there is any growth then a sensitivity disc is placed on a pure culture of isolate.
Gram staining is then carried out on each specimen, a wet preparation of the swab can be made by dropping normal saline into the swab container and the swab stick is rubbed on a slide covered with a slip and viewed under the microscope; pus cells, epithelial cells, yeast cells etc can be viewed.
Result: Possible pathogens include; Neisseria gonorrhea, Trichomonas vaginals, Candida spp, Clostridium perfrigens etc.
3.6.5 MANTOUX SKIN TEST
Introduction: The Mantoux test or Mendel-Mantoux test (also known as the tuberculin sensitivity test, or PPD test for purified protein derivative) is a screening tool for tuberculosis (TB) It is one of the major tuberculin skin tests used around the world, largely replacing multiple-puncture tests such as the Tine test. Tuberculin is a glycerol extract of the tubercle bacillus. Purified protein derivative (PPD) tuberculin is a precipitate of species-nonspecific molecules obtained from filtrates of sterilized, concentrated cultures.
Material/Equipments: Millimeter rule, tuberculin injection, Personal protective equipment, cotton wool.
Procedures: A standard dose is 5 tuberculin units (TU - 0.1 ml) is injected intradermally (between the layers of dermis) and read 48 to 72 hours later. This intradermal injection is termed the Mantoux technique. A person who has been exposed to the bacteria is expected to mount an immune response in the skin containing the bacterial proteins. If a person has had a history of a positive tuberculin skin test, or had a recent tuberculin skin test (within one year), another skin test should be used
Result: Within two days of injection, the reaction is read by measuring the diameter of induration (palpable raised, hardened area) across the forearm (perpendicular to the long axis) in millimeters. If there is no induration, the result should be recorded as 0mm. Erythema (redness) should not be measured. A tuberculin test conversion is defined as an increase of 10 mm or more within a two-day period, regardless of age. Alternative criteria include increases of 6, 12, 15 or 18mm.
CHAPTER FOUR
4.0 SUMMARY, CHALLENGES ENCOUNTERED, AND CONCLUSION RECOMMENDATION
4.1 SUMMARY OF ATTACHMENT ACTIVITIES
During my period at the Orile Agege General Hospital as a SIWES student, cataloguing some information materials for the laboratory and I also did some activities at the reception such as: attending to patients, confirming and examining their request forms, entering their details into the register, detailing them concerning the test they are to undergo and directing them to where is to be carried out.
I was later transferred to the laboratory and was introduced to the departments, safety precautions and tests carried out in each department.
4.2 CHALLENGES ENCOUNTERED
The main problems encountered were getting placement and transportation. It was quite challenging for me that live in far place to get to the organisation every working day. I was not given any remuneration or allowance, other problems encountered during the training was attending to different people with different personalities at the reception.
4.3 CONCLUSION
My four months industrial attachment with Orile Agege General Hospital has been one of the most interesting , productive, instructive and educative experience in my life. Through this training, I have gained new insight and more comprehensive understanding about the real industrial working condition and practice and also improved my soft and functional skills.
All these valuable experiences and knowledge that I have gained were not only acquired through the direct involvement in task but also through other aspects of the training such as: work observation, supervision, interaction with colleagues, supervisors, superior and other people related to the field. It also exposed me to some certain things about medical environment. And from what I have undergone, I am sure that the industrial training programme has achieved its primary objective.
As a result of the programme, I am now more confident to build my future career which I have already started with Orile Agege General Hospital.
4.4 RECOMMENDATION
I recommend that all institutions or bodies involve in Student Industrial Working Experience Scheme, should provide places for industrial attachment for Student Industrial Training Fund and also pay some allowances to students and the company should provide more safety equipments to prevent further environmental and health hazards.
Also, to students that are to undergo the training, I recommend that they should take it very seriously, because it is one of the most important parts of their studies which will help them build a very significant and effective meaning in their career pursuit.
