CHAPTER ONE

1.1 Introduction.
Students industrial works experience scheme (SIWES) was established by the federal
government of Nigeria in October 8, 1973, with it headquarter located in Miango, Plateau State.

The SIWES is the accepted technical training program which forms an integral part of the
approved minimum Academic standards in the various degree programs for Nigerian
universities.

It is aimed at exposing students to instruments and equipment, professional work methods and
ways of safe guarding the work areas and workers in Industries. The effort is aimed at helping
the students in the Nigerian tertiary institutions to put into practice the theoretical aspect of their
studies field.

1.2 Aim and objective of SIWES.

The following are someof the aims and objectives of SIWES.

(a) It create avenue for students in higher institutions of learning to put into practice what has
been learnt at school.

(b) It gives students the opportunity to apply their knowledge gained practically.

C. It prepares students for graduate work.

Benefits of the scheme

i. Bridge the gap between the theories learned in the class and the practicals that are found
in the field or industries
ii. Consolidation of class room studies with methods of working and experience in
traditional and machines in industries.
iii. Prepareds students for business careers by merging analytical experience with self-
relience.
iv. Provides opportunity for additional research by students.
1.3 Historical background of the organization.
Christian Reformed Church Nigeria (CRCN) comprehensive health center and maternity wukari
is a faith based clinic. It is a product of CRCN Rural Health Program initiative in a bid for
Wholistic ministry. The establishment dated as far back as 1953. More proactive work started in

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1970, with the main objective of bringing medical services to rural areas, using medicine as a
doorway to evangelism, regardless of tribal or religion affiliation, age or financial status.

It also partner with Integrated Development Program (IDP), Global Fund, USAID, Institute of
Human Virology of Nigeria (IHVN) and National Action Committee on AIDS (NACA) in
providing intensive health care services for HIV/AIDS patients.

1.4 Objectives of the organization:

i. To engage in activities that promote better health care for community.
ii. To propagate the gospel of the Lord Jesus Christ.
iii. To have a doctor on seat that provides surgery and specialized treatment.
iv. To carry out immunizations as required.
v. To maintain a viable drug store that is current.
vi. To procure facilities that will provide care.
1.5 Organizational chart

MANAGING DIRECTOR

ACCOUNTANT

CLINIC INCHARGE

MATERNITY ASST. CLINIC LABORATORY

INCHARGE INCHARGE INCHARGE PHARMACIST

METRON BIRTH
NURSE I NURSE II
ASST. LAB
MOTHER ATTENDANT
INCHARGE

HEALTH
RECORD
CLINIC
ATTENDANT
COMMUNITY LAB
VOLUNTEER
ATTENDANT

CASHIER CLEANER SECURITY

Figure 1: Organisational chart

1.6 Different sections/ units of the laboratory.

CRCN comprehensive health center and maternity wukari is divided into departments
and sections, some of which is subdivided in to other units. The Laboratory department is
divided into the following units:
1. Central collection unit:

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 2. Clinical chemistry (Biochemistry): This unit commonly called chemical pathology
includes instrumental analysis of blood components, enzymology, toxicology and
endocrinology.
3. Clinical microbiology: This section encompasses three different units. These include.
(a) Bacteriology
(b) Serology
(c) Parasitology.
4. Haematology and Blood group serology (BGS): This unit consists of automated and
manual analysis of blood cells.

1.7 Safety rules in Medical Laboratory

i. Clean the work bench before and after work.
ii. Always put on laboratory coat and hand gloves while working in the laboratory.
iii. Always take every sample in the laboratory to be pathogenic.
iv. In case of accident with chemicals ensure to clean the place with clean water and
detergent and report to the laboratory incharge.
v. Always wash hand before leaving the laboratory.
vi. Always label samples immediately after collection.
vii. Switch off electrical appliances after use and before leaving the laboratory.

1.8 Equipment and apparatus used in the laboratory.

i. Sample collection vials: These are tubes containing different kinds of anticoagulant
used for collecting patients’ blood samples.
ii. Graduated pipette: This is used in pipetting samples and other Liquids to be used in
investigations.
iii. Automatedpipette: The automated pipette is used for pipetting samples in microliters.
iv. Test tubes: Test tubes are used in putting sample for investigation.
v. Test tube rack: It is used for holding test tube in position.
vi. Triple beam balance: It is used to measure solid substances.
vii. Centrifuge: It is used for spinning samples to fractionate its components such as cells,
which may be suspended in a fluid. The principle is that the centrifuge exerts a

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 centrifugal force which is greater than that of gravity, and causes particles in a fluid to
sediment.
viii. Refrigerator: The refrigerator is used in storing samples and reagents that need to be
kept at low temperature.
ix. Microscope: It is used in viewing objects that are not visible to the naked eye.
x. Spectrophotometer: It is an instrument used in reading the absorbance (optical
density) of liquid samples.
xi. Hematocrit machine: It is used for carrying out packed cell volume.
xii. ReflotronDry chemistry analyzer: it is used for carrying different chemical analysis.
xiii. Abacus 380 Hematology analyzer: it is used for carrying out full blood count.
xiv. BD FACS count: it is used for estimation CD4 cells in HIV patients.

Figure 2.1 : Microscope

Figure 2.2: Abacus 380 Haematology Analyzer

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Figure 2.3: Reflotron chemistry Analyzer

Figure 2.4: Photospectrometer

Figure 2.5: Tripple Beam balance

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Figure 2.6: BD Facs Count Machine

Figure 2.7: Glucometer

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 CHAPTER TWO

2.1 Major Activities Carried Out In the Laboratory

The followings are the major activities carried out in the laboratory

(a) Liver function test (LFT)

(b) Fasting/Random blood sugar (FBS)
(c) Cholesterol.
(d) Urinalysis
(e) Pregnancy test (PT)
(f) Hepatitis Rapid test.
(g) HIV/AIDS
(h) Microscopy (urine & stool)
(i) Malaria parasite (MP)
(j) Widal test (typhoid fever)
(k) Semen analysis (SFA).
(l) Blood Grouping and cross matching
(m) CD4 count
(n) Haemoglobin Estimation
(o) Erythrocyte Sedimentation Rate (ESR)
(p) Full Blood Count
(q) Packed Cell Volume

2.2.0 Description Of Each Investigation Conducted

2.2.1 Sample Collections
Procedure for vein-puncture blood collection:

(i) The tourniquet was tied at the upper arm 5cm from the elbow.
(ii) The alcohol swap was used to disinfect the area of the skin with prominent vein.
(iii) The needle was gently inserted horizontally into the vein and blood was drawn. The
tourniquet was untied and the needle was gently removed.

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 (iv) The blood sample was dispensed into a centrifuge tube containing anticoagulant
(fluoride oxalate) for glucose estimation or EDTA for other hematological test) and
centrifuge at 2000rpm for 3mins.
(v) The sample was separated using dry Pasteur pipette into a dry chemically clean plain
container and was labeled.

Procedure for urine collection.

Urine sample is collected in a clean dry container. Usually, the first part of the urine is
allowed to flow out and the middle part is collected in the clean dry sterile container.

Procedure for stool collection

Stool samples are collected in a clean dry container

Procedure for semen collection.

The sample is collected after 2-3 days abstinence from sexual intercourse or preferably by
masturbation.Thecollected sample is delivered within two hours of collection.

Procedure for urethralswap collection.

i. The wire loop was sterilized and allowed to cool for a few seconds.
ii. The wire was then inserted into the penis through the opening to collect the fluid.
iii. The swap was collected and cultured immediately.

2.5.0 Lipids and Lipoprotein

2.5.1Estimation of serum/plasma cholesterol. (Manual method)
Aim: To estimate serum cholesterol.

Principle: In the presence of excess acid such as phosphoric acid and ferric (fe3+) ions,
cholesterol is oxidized to disulphonic and which is reddish purple in color. It is read
spectrophotometrically at 560mm against standard and a blank.

Reagents: ferric chloride reagents, cholesterol standard, serum/plasma (after 12hrs of fasting).

Procedure for test:

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 Test Standard Blank

Ferric chloride 5.0ml 5.0ml 5.0ml

reagent

Diluted serving 0.5ml - -

Diluted standard - 0.5ml -

Diluted water - - 0.5ml

The above table shows the procedure for carrying out cholesterol estimation.

i. The serum was diluted 1:20 with distilled water (0.1m1 of serum + 1.9ml of distilled
water).
ii. The cholesterol standard was diluted at 1:20 with glacial acetic acid.
iii. Three (3) tubes was set up as shown in the below table.
iv. Each test tube was shake for 10secs to mix the content
v. The test tubes were immediately place in a booking water bath for 90secs.
vi. It was allowed to cool in running tap water for 5mins.
vii. The absorbance was read at 560mm against the blank and standard.

Calculation:

|of |test
Serum cholesterol (mg/dl) = ×250
|of |standard

Serum cholesterol (mol 11) = mg/dl 0.0259

Normal values. For adult: from 2.5 to 6.3mol/l.

Estimation of serum/plasma cholesterol. Automated

Title: Quantitative determination of cholesterol in blood/serum/plasma

Aim: To determine quantity of cholesterol in blood/serum/plasma

Introduction: Cholesterol is a steroid with a secondary hydroxyl group in the C3 position. It is

synthesized in many tissues, but particularly in the liver and the wall of the intestine. About three

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quarter of the body’s cholesterol is synthesized by the tissues while one quarter comes from the
diet. Determination of cholesterol is used to screen for an atherogenic risk and diagnosis and
treatment of disease with elevated cholesterol level and disorders of lipid and lipoprotein
metabolism.

Principle:After application the the test strip, the sample flows into the reaction zone, where the
cholesterol esters are cleaved into the corresponding fatty acid and cholesterol which are then
oxidized to cholesterone and hydrogen peroxide in the presence of oxygen. In a further reaction
step catalyzed by the enzyme peroxidase, the hydrogen peroxidase oxidezed a redox indicator,
resulting in a blue dye which is proportional to the cholesterol concentration in the sample.

Cholesterol + ester + H2O-------Chol esterase--------- Cholesterol + RCOOH

Cholesterol + O2-----------------chol esterase-------------Choresterone + H2O2

H2O2 + indicator -----------------peroxidase-------------- Blue dye + H2O

Procedure:
i. the dry chemistry analyzer was Switched on
ii. When the display shows READY, CHOL test strip was removed from the container
and unwrapped.
iii. Using the pipette, 32 µl of the sample was drawn and dispensed on the application
zone.
iv. The sliding cover was opened and the strip was inserted and the sliding cover was
closed back.

Result
Normal Value < 200mg/dl or <5.2mmol/dl

2.5.2urine analysis.
Title: Qualitative analysis of urine
Aim: To determine cause of urogenital track infection
Introduction: The physical, chemical and microscopic examination of urine is known as urine
analysis or routine urinalysis. Urinalysis is an important part of the initial examination of patient

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and the result provide a valuable picture or information of the patient’s general health pattern.
General urine analysis indicates the following:

i. The state of the kidney and the urinary tract.

ii. Information about metabolic and systematic (non-renal) abnormalities.

Collection and preservation of urine sample for urinalysis

Urine sample for urinalysis is collected in a clean dry container and examined as

Soon as possible. In most cases, the first specimen freely voided in the morning is preferred.
Although, specimen collected 2-3hrs after eating is more suitable when testing for glucose. On
standing, red blood cells and white blood cells are destroyed; casts are decomposed; glucose is
lost and Bacterial contamination of the urine takes place causing alkalization of the sample.
Therefore, the specimen must be examining when fresh, ideally within 30mins of collection.

Physical properties Color,Transparency,Odor, Specific

gravity(urinometer)

Physical screening test(reagent strip) PH, Specific gravity, Protein, Blood, Glucose,
Ketone, Bilirubin, Urobilirubin.

Microscopic examination of urine sediment Red blood cells, White blood cells, Epithelial
cells, cast, Bacterial and other Microorganisms,
Crystal, Other components.

Table 2.0

The above table shows different component in the urine.

Physical examination of urine:

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Volume: Under normal condition there is a correlation between urine volume and water intake of
a person. Certain conditions however give rise to increase or decrease in the volume of urine.
Some of these conditions are listed below:

Polyuria: Is an excessive or an abnormal large production or passage of urine (greater than 2.5
or 3L over 24hrs in adult)

Oliguria: Decreased in the volume of urine (<200ml/24hrs).

Anuria: Complete suppression of urine formation.

Nocturia: Excess urine at night (>500ml).

Color: The color of normal urine varies greatly even in one person in a single day. In general,
the normal urine color is described as straw, yellow or amber. This normal color is due to three
(3) pigments: urochrome, uroerythin and urobilin.

Transparency: Normal urine is clear when freshly voided but it often becomes cloudy when
allowed to stand for some time. Cloudiness is due to precipitation of chemical substance (crystal,
amorphous phosphate, and urate) in urine or due to bacterial growth.

Odor: Freshly voided urine has a faintly aromatic odor due to the presence of certain volatile
acid. Bacterial action occurs when urine is allowed to stand and this produces ammoniacal odor.

Foam: Normal urine will produce a moderate amount of white foam when shaken. A urine
sample with high protein concentration will produce large amount of foam when shaken.

Specific gravity: Specific gravity (sp.gr) of urine is the measure of the amount of dissolved
substance in the urine. It is the weight of the urine compared to the weight of an equal volume of
distilled water at constant temperature. Urine of low sp.gr. Is called hyposthenuric (˂1.007) and
those of fixed specific gravity are isothenuric (abet1.010).

weight of urine
Specific gravity =
weigh of equal vol . of distilled water

2.7 .0 Endocrine Function Test

2.7.1Pregnancy test:
Title: Determination of human chorionic gonadotropin in urine

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Aim: To carry out pregnancy test using HCG(human chorionic gonadotropin) strip.

Introduction: The HCG one strip (urine or plasma) is a rapid chromatographic immunoassay for
qualitative detection of human chorionic gonadotropin in urine to aid in the early detection of
pregnancy.

Principle: The HCG one step pregnancy test uses two lines to indicate result. The test utilizes a
combination of antibodies including a monoclonal HCG antibody to selectively detect elevated
level of HCG. The control line is composed of coat polyclonal antibodies and colloidal gold
particles.

Procedure for test:

i. The sealed pouch was removed by tearing the notch.

ii. The test strip was immersed into the sample, not exceeding the MAX line with the
arrow pointing toward the sample.
iii. It was allowed for 15mins for color band to develop.

Result.

Two lines appear at the control and test region = positive (+)

One line appear at the control region only = Negative (-)

Precautions

i. The test strip was used before its expiration date.

ii. The test strip was not allowed to stay out of its pouch for so long after it was removed

before used.

iii. All the specimens were considered as potentially hazardous and so treated as such.

iv. The test strip was discarded in a proper biohazard container after testing.

2.8.1Widal test (for typhoid fever), the slide method

Title: Determination of Salmonella Species using Widal method

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Aim: to determine the presence of Salmonella species in Serum/plasma
Introduction: Enteric fever occur when pathogenic microorganism like salmonella typhi or
paratyphi infect the human body during the course of disease the body responds to this antigenic
stimulus by producing antibodies whose titre rise slowly in early stage to a maxima and the
slowly fails till it is undetectable. The sub species enterica with serotypes salmonella
Typhimurium.
The serotypes are routinely identified because of their clinical significance. They are as follows:-
Salmonella paratyphi A (Sero group A)
Salmonella paratyphi B (Sero group B)
Salmonella choleraesuis (Sero group C)
Salmonella typhi (Sero group D)
The major antigen in salmonella typhi are:
i. Somatic antigen “O”
ii. Flagella antigen “H”
Principle: when the coloured smooth attenuated Widal test antigen suspensions are mixed with
the patient serum, antisalmonella antibodies present in the serum react with the antigen
suspension to give agglutination.

Materials: White Rocking Tile, WIDAL Kit, test sample (Serum).

Test procedure:

a. The test sample was placed on the white tile forming 8 wells (4 wells in two rows).
b. The reagent from the WIDAL Kit wasadded to the sample, the O antigen to the first row and
the H antigen to the second row.
c. The tile was rocked for 30 seconds to 1minutes and observed for agglutination.
d. The result was recorded based on the significant titer of 160

Result:

The agglutination are scored as follows: 1/20, 1/60, 1/80, 1/120, 1/160, 1/320

Precaution:

i. The result was read between 30 - 60 seconds

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 ii. The slide was thoroughly clean before use.

Syphilis(VDRL) strip test

Title: Determination of the presence of Treponema pallidum
Aim: To determine the presence of Treponema pallidum
Introduction: The test is conducted to detect the presence of bacterial treponema pallidum (T.
pallidum) a sexual transmitted infection (STI) known as syphilis.
Principle:One step Anti-TP in whole blood rapid screening test is a chromatographic
immunoassay for the qualitative detection of Antibodies of TP in human whole blood. Two
purified recombinant antigen of TP are used in the test band as captured material and gold
conjugates. If the antibodies of Anti-TP is present in the sample in concentration above the
labeled, complex will be formed. This complex is then captured by antigens immobilized in the
test zone of the membrane, producing a visible pink-rose color band on the membrane. The color
intensity depends on the concentration of Anti-TP present in the sample. This one step test is
very sensitive and it only takes about 15minutes. Test results are read visually without an
instrument.
Test procedure:

a. The sealed pouch was removed by tearing along the notch

b. The test strip was immersed into the sample not past the MAX line with the arrow pointing
towards the sample.

c. It was allowed for 15minutes for color band to develop.

Result:

Two lines appear at the control and test region=POSITIVE

One line appear at the control region only =NEGATIVE

Precautions:

i. The test strip expiration date was checked before use

ii. The result was read between 30 – 60 seconds

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2.9.0.0Virology
2.9.0.1HIV/AIDS related complex [retro viral screening]
Title: determination of HIV using rapid test
Aim: to determine the presence of human immunodeficiency virus antibodies in the serum.
Introduction: AIDS (acquired immunodeficiency syndrome) is characterized by changes in the
population of T-cell lymphocyte. In an infected individual, the virus causes depletion of helper
T-cells, which leaves the person susceptible to opportunistic infections and some malignancies.
The virus that causes AIDS exists as two related types known as HIV-1 and
HIV-2. The presence of the aids virus triggers the production of specific antibodies to either
HIV-1or HIV- 2.

Principle: Alere determiner HIV-1/2 is an immunochromatographic test for the qualitative

detection of antibodies to HIV1/2. A sample is added to sample pad. As they migrate through
the conjugate pad, it reconstitutes and mixes with selenium colloid-antigen conjugate. This
mixture continues to migrate through the solid phase to the immobilized recombinant antigens
and synthetic peptides at the patient window sites. If antibodies to HIV-1 and /or HIV-2 are
present in the sample, the antigen selenium colloid flows past the patient window and no red line
is formed in the patient window site.

Test procedure

1. The protective foil cover was removed.

2. 2-3 ml of the blood sample (serum) was pipette using a Pasteur pipette and drop in the
sample pad.
3. It was allowed to stand for 15minutes and the result was read.

Interpretation of result

i. Two red bars appear in both the control and the patient window =POSITVE.
ii. One red bar appears in the control window of the strip =NEGATIVE.
iii. No red bar appears on both control and patient window=INVALID.

2.9.1.0 Parasitology
2.9.1.1Malaria Parasite
Title: determination of malaria parasites
Aim: To determine the presence of malaria parasite in human blood.

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Introduction: MP test is carried out to identify the presence of parasite that causes
plasmodiasis.The genus plasmodium (P) are divided to four (4) species (SPP) which are:

i. Plasmodium ovale
ii. Plasmodium malariae
iii. Plasmodium vivax
iv. Plasmodium falciparum
The P. falciparum exists in the tropic and sub-tropic and is responsible for approximately 50% of
all malaria cases P. falciparaium usually only young trophozoites and gametocytes are seen in
peripheral blood smear.
To carry out this test, microscopic mode of identification and slide stained preparation is used.
Materials used: microscope slide, microscope, lancet, field stain A and field stain B, staining
rack.

Test procedure:

i.The tip of the finger was pricked with the lancet.

ii.Blood from finger was smeared on the slide to make a thick film.

iii.The slide was allowed to air dry.

iv.The slide was then stained with the field strain A for 1minute and washed off, then stained
again with field stain B and washed of immediately.

v.The slide was allowed to air dry.

vi. Immersion oil was added and the sample viewed for the presence of malaria parasite under
the x100 objective lens.

Result:

1-10 parasites in 100HPF (High Power Field) +

Is reported as trophozoite of P. falciparum 1+ seen

More than 10 parasite in 100 HPF ++

Is reported as trophozoite of P. falciparum 2+ seen

Between 1 – more than 10 in each 100 HPF +++

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 Is reported as trophozoite of P. falciparum 3+ seen

Table 3.1

Precautions:

i. The microscope objectives and slide were clean with sterile tissue.
ii. The stains were applied appropriately.

2.9.3.0 Blood grouping.

Title: Determination of blood group using the direct slide method.

Aim:To determine blood group (ABO system).

Principle: If the red blood cell contains the corresponding antigen and antibody, there will be a
reaction which results to agglutination (Clumping) of the red cells. But if either antigen or
antibodies areabsent, there will be no agglutination.

Reagents: ABO grouping antisera (Anti-A, Anti-B and Anti D)and whole blood sample, rocking
tile, mixing rod.

Procedure:

a. A clean white tile was marked with three wells; a drop of blood was placed in each of the
well.
b. A drop of Anti-A was added to the first well, a drop of Anti-B was added to the second
well and a drop of Anti-D was added to the third well.
c. The tile was rocked gently for 2mins.
d. The tile was observed and the presence or absence of agglutination was noted.

Type A+ A- B+ B- AB+ AB- O+ O-

Anti-A + + - - + + - -

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 Anti-B - - + + + + - -

Anti-D + - + - + - + -

Result

Table 4.1
The table above shows the results of blood grouping.

Note: (+) means agglutination is seen while (-) means that no agglutination seen

Blood group compatibility chart

RECIPIENT GROUP

X A+ A- B+ B- AB+ AB- O+ O-

A+ √ X X X √ X x X

A- √ √ X X √ √ x X

B+ X X √ X √ X x X

B- X X √ √ √ √ x X

AB+ X X X X √ X x X

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 D AB- X X X X √ √ x X
O
O+ √ X √ X √ X √ X
N
-
OO √ √ √ √ √ √ √ √

G
R
O
U
P

Table 4.2Key: (√) means compatible (x) means not compatible

Precautions:

i. The anti-sera were stored at appropriate temperature

ii. The result was read between 1-2 minutes

2.9.3.1Hematocrit or packed cell volume (PCV).

Title: determination of packed cell volume of a given sample.

Aim: to determine a packed cell volume of a given sample.

Introduction: Hematocrit or packed cell volume is a percentage (%) of the total volume of a
whole blood occupied red blood cells. When a whole blood is spin or centrifuge at a constant
speed at a constant period of time.

Principle:The principle works based on the fact that, when a venous red blood cells is filled in a
heparin pre-coated capillary tube of about 7cm in length and 1mm in internal diameter and
centrifuged at a certain revolution for a certain period of time, the red blood cells settles at the
base of the tube, while the serum is suspended at the top of the capillary tube. The result is read
in percentage (%).

Procedure:
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 a. The capillary tube was filled with a well-mixed venous blood (or a direct puncture of the
finger tip).
b. The filled capillary tube was placed in a micro-hematocrit centrifuge and spin at 12,000g
for 5mins
c. The spincapillary tube was placed into an aseptically designed scale and the result of the
PCV was read and expressed inpercentage (%).

NOTE: In the absence of the specially designed scale, PCV can be calculated as shown below:

packed RBC column height

PCV % ¿ ×100
total blood column height

Normal range: Men: 40-52% (0.4-0.52L/L)

Women: 37-47% (0.37-0.47L/L).

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