CHAPTER ONE

INTRODUCTION

Student Industrial Work Experience Scheme (SIWES) is a very big aid

and a stepping stone to life after school. It is an opportunity given to students to

put into practice most things that were theoretically explained by lecturers in

schools.

Pathology laboratory in General Hospital Onitsha, Anambra State has

been of great aid to this programme most especially for science students due to

the fact that they are opportune to make use of various reagents and equipments

which may not be available to them in their schools.

As a result of this, the standard methodology and equipment handling

expose students in their course of training.

1.1 A. Meaning of SIWES

Student Industrial Work Experience Scheme, SIWES, is the accepted skill

training programme which forms part of the approved minimum academic

standards in the various degree programmes for all the Nigerian universities. It

is provided to bridge the gap existing between theory and practice of

engineering, science and technology, agriculture, medicine, management and

other professional educational programmes in the tertiary institutions.

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 It is aimed at exposing students to machines and equipments, professional

work methods and ways of safe-guarding the work areas and workers in

industries and other organizations.

B. Objectives of SIWES

1. To prepare students for work situations they are likely to meet after

graduation.

2. To provide an avenue for students in Nigerian universities to acquire

industrial skills and experience in their course of study.

3. To enlist and strengthens employers involvement in the entire educational

process of preparing university graduates for employment in industries.

4. To provide students with an opportunity to apply their theoretical knowledge

in real work situations, thereby closing the gap between university work and

actual practice.

5. To expose students to work methods and techniques in handling equipments

and machinery that may not be available in the universities.

6. To make the transitions from the university to the world of work easier and

thus enhance students contact for later job placement.

7. Teaches the student on how to interact effectively with other workers and

supervisors under various conditions in the organization.

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1.2 History of General Hospital Onitsha, Anambra State

Onitsha General Hospital was established in 1999 by the Anambra State

Government to enhance and promote the health services render by the Anambra

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

Anambra State.

OPERATIONS OF THE ORGANIZATION

The organization is a Hospital and Maternity center with a wide range of

medical staff capacity which includes: Gynecologists, Pediatricians,

Dentists, Opticians, Consultants, General medical Practitioners, Nurses,

laboratory scientists and technicians, etc. There are other major areas of

operation. These areas include;

 IMAGING: This is one of the sensitive departments of the

organization. It includes Mammogram, Three and Four- Dimensional

Forms (3D & 4D) colored ultrasound Scan and Digital X-RAY.

 PHARMACY: This department is headed by a head Pharmacist, who

oversees the affairs of the Pharmacy. Here, we find a wide variety of

drugs ranging from Anti-biotics, Anti-fungal, Anti-malaria and

Creams/Ointments etc.

 DIAGOSTIC LABORATORY: It is headed by a Pathologist, who

oversees the affairs of the Laboratory scientists, Technicians and

Phlebotomists.
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 Other departments found in the organization ranges from The

Administrative Department, Accounting, Front Desk Office, Public Relations

Office, Health Maintenance Organization Office, Billing Office, Cash Office,

Janitorial, Transportation, Laundry, Gardening, Security, Kitchen, etc.

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 1.5 ORGANISATION’S CHART AND ORGANOGRAM

Onitsha General Hospital

Chief medical director

Hospital management board Hospital Administrator

Finance Office Records Hospital Matron Admin Officer 11

Gynecology Obstetrics Physiotherapy X-ray

Ddtj Laboratory Pharmacy Nursing Security

Maternity

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 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.

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 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.

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  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.

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  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.

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 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.

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 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).

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 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

3.1.1IDENTIFICATION OF MATERIALS USED IN THE PHLEBOTOMY AND

USES

1. Personal Protective Equipment

 Phlebotomy Uniform: Serve as protective to the body

 Disposable gloves/Hand Gel: For protection against spillage

2. Needles: Used to make incision

3. Needle holders: For holding needle or also known as venoject

4. Vacutainers, Vacutainer holder or Syringe: Serves as blood drawer

5. 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.

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 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.

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 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.

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6. 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.

7. 70% Alcohol swabs: Used as a disinfectant for the area in which incision is to

be made.
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 8. Micropore tape: To aid hemeostatis

9. Dental rolls

10. Adhesive dressing

11. Racks

12. Disposable waste bins

13. Cotton wool Pillow or other support: Used for supporting the arm for easy

blood flow

3.1.2STANDARD 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.

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 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.

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 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.

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 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.

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3.1.3 PHLEBOTOMY DIAGRAMS

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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.

3.2.1MATERIALS 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.

3.2.2COMPLETE BLOOD COUNT (CBC) TEST

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 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.

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  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

3.2.3PACKED 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.

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 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

sample2:and
PLATE One end
MICRO of the tube was
HAEMATOCRIT sealed with flame using the Bunsen burner,
READER

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

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 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

3.2.4ERYTHROCYTE 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.

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 Equipment/Apparatus: Sodium Citrate bottle, ESR rack, Westergren glass rod.

26
  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

3.2.5BLOOD 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

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 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

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 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.

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30
  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

3.3.1HBs.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

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 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

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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

33
 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

3.3.3WIDAL 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

34
 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)
35
 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.

36
  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.

3.4.1MATERIALS 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.

3.4.2BLOOD GLUCOSE
37
 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.

1. 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

38
2. 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.

3. 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

4. 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

39
 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

5. 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

40
 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

3.4.3ELECTROLYTES 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

3.4.5BLOOD UREA NITROGEN AND CREATININE TEST

41
 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

3.4.6LIVER 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
42
 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

3.5.1BASIC 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.

3.5.2GENERAL 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.

43
 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.

44
3.5.2MATERIALS/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.

3.5.3INTRODUCTION 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,

45
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.

46
 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.

3.5.4COMMON MEDIA IN ROUTINE USE

1. 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

2. 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:

a) Selective: as bile salt does not inhibit the growth of enterobactericeae but

inhibits growth of many other bacteria.

b) 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,

47
 then boiled for one minute to completely dissolve the powder. The Agar was

autoclaved at 121OC for 15minutes.

3. 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

4. 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,

5. 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

48
 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.

3.5.6PRECAUTIONS 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.

3.5.7GRAM STAINING

49
 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
50
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.

51
3.5.8PROCEDURE 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 121⁰C for 15minutes. The safety valve was closed at 120⁰C and

at 121⁰C, button was switched off. The medium were allowed to cool for 47⁰C

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 121⁰C is referred

to as culture media autoclave.

52
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.

3.6.1STOOL 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

53
 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;

1. Macroscopic-----------------Hard, dark ,brown stool formed with no mucus or

blood seen.

2. Microscopic-----------------Present of Ascaris lumbricoides .

3.6.2URINE 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.

54
 Culture: A loopful of the sample was used to make a streak on the Agar plates.

The plates were then incubated for 24hours at 37 OC. 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

1. Appearance: Clear, turbid ,slightly turbid,

2. 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

3.6.3EYE, 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;
55
  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

3.6.4HIGH 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.

56
  Result: Possible pathogens include; Neisseria gonorrhea, Trichomonas vaginals,

Candida spp, Clostridium perfrigens etc.

3.6.5MANTOUX 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

57
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.

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 CHAPTER FOUR

4.0 SUMMARY, CHALLENGES ENCOUNTERED, AND CONCLUSION

RECOMMENDATION

4.1 SUMMARY OF ATTACHMENT ACTIVITIES

During my period at the Onitsha 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.

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 CHAPTER FIVE

CONCLUSION AND RECOMMENDATION

5.1 CONCLUSION

My four months industrial attachment with Onitsha 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 Onitsha General Hospital.

5.2 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

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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.

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