Augustine DuduBabe medical Laboratory

25/09/2025

Semen Analysis
1. Objective
The objective of the semen analysis was to evaluate the quality and fertility potential of a male semen sample by examining its physical, chemical, and microscopic characteristics.
2. Principle
The principle is that semen quality could be determined by assessing physical parameters (volume, color, viscosity, pH), motility of s***matozoa, concentration (s***m count), morphology, and the presence of abnormal cells or contaminants. These parameters reflected the functional capacity of the male reproductive system.
3. Materials
• Sterile wide-mouth semen collection container
• Microscope (light microscope with phase-contrast if available)
• Glass slides and coverslips
• Counting chamber (e.g., Neubauer chamber)
• pH paper or pH meter
• Timer/stopwatch
• Personal protective equipment (gloves, lab coat, mask)
• Incubator (if needed for liquefaction observation)
4. Procedure
1. The semen sample is collected after 3–5 days of abstinence, by ma********on into a sterile container.
2. The sample is allowed to liquefy at room temperature for 30 minutes.
3. Physical examination was performed, noting volume, color, viscosity, and pH.
4. A drop of semen was placed on a slide, covered, and observed under the microscope for motility (progressive, non-progressive, immotile).
5. The s***m concentration was determined using a Neubauer chamber.
6. Smears is prepared and stained for assessing s***m morphology.
7. The presence of pus cells, red blood cells, or epithelial cells was also recorded.
5. Result
• Normal findings (WHO reference values):
# Volume: ≥1.5 mL
# pH: 7.2–8.0
# S***m count: ≥15 million/mL
# Motility: ≥40% motile
# Morphology: ≥4% normal forms
• Abnormal findings included low volume, low s***m concentration (oligos***mia), absent s***m (azoos***mia), poor motility (asthenozoos***mia), or abnormal morphology (teratozoos***mia).
6. Uses
• It is use for male infertility evaluation.
• It helped monitor the success of vasectomy or other rep

24/07/2025

HBsAg Test (Hepatitis B Surface Antigen Test)
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∆. Objective:
The objective of the HBsAg test is to detect the presence of Hepatitis B surface antigen in a patient’s blood, which indicated an active Hepatitis B virus (HBV) infection.
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∆. Principle:
The test is based on immunoassay techniques, such as ELISA or rapid chromatographic methods. It utilized antibodies that specifically bound to the HBsAg if present in the sample, producing a visible signal or color change.
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∆. Materials:
• Patient's serum or plasma sample
• HBsAg rapid test cassette or ELISA kit
• Sample dropper or micropipette
• Buffer solution
• Gloves and lancet (for sample collection)
• Timer
• Biohazard disposal container
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∆. Procedure (Rapid Test):
1. A blood sample is collected from the patient.
2. 2–3 drops of the serum or plasma were added to the sample well of the test cassette.
3. 1–2 drops of buffer were added.
4. The test was allowed to run for 15–20 minutes.
5. The result is read visually by observing the appearance of colored lines.
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∆. Result:
• Positive: Two colored lines appeared — one in the control (C) region and one in the test (T) region, indicating the presence of HBsAg.
• Negative: Only one colored line appeared in the control (C) region, and no line appeared in the test (T) region.
• Invalid: No line appeared in the control region — the test is considered invalid and needed to be repeated.
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∆. Uses:
• It is used to screen for Hepatitis B virus infection.
• It helped identify both acute and chronic HBV infections.
• It was part of routine health screenings, blood donor screening, and prenatal checkups.
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∆. Consultation:
Patients with a positive result are to be advised to consult a healthcare provider or hepatologist. Further confirmatory tests such as HBeAg, anti-HBc, and HBV DNA were recommended. Counseling regarding transmission prevention, vaccination of close contacts, and possible antiviral treatment was also provided.

Thanks for your Time ••••••••••👏

TUT.LT.Aryamu Augustine Lupo 🇸🇸

24/07/2025

HCV Antibody Test
✓. Objective:
The objective of the HCV antibody test is to detect antibodies against the Hepatitis C virus in a patient's blood, indicating past or present infection.
✓. Principle:
The test is based on immunochromatographic or ELISA methods. It used antigens specific to HCV that bound to anti-HCV antibodies in the patient's sample. A color change or line on the test device indicated the presence of antibodies.
✓. Materials:
• Patient’s blood, serum, or plasma
• HCV rapid test cassette or ELISA kit
• Buffer solution
• Micropipette or dropper
• Lancet and gloves
• Timer
• Alcohol swab
• Biohazard disposal container

✓. Procedure (Rapid Test):
1. A blood sample is collected through finger prick or venipuncture.
2. 2–3 drops of the sample are placed into the test cassette’s sample well.
3. 1–2 drops of buffer were added.
4. The test cassette was left undisturbed for 15–20 minutes.
5. The result is read by checking the appearance of colored lines.

✓. Result:
• Positive: Two lines appeared — one in the control (C) region and one in the test (T) region, indicating anti-HCV antibodies were present.
• Negative: Only one line appeared in the control region; no line in the test region.
• Invalid: No control line appeared — test result was invalid and had to be repeated.
✓. Uses:
• It is used to screen for HCV infection in at-risk populations.
• It aided in diagnosis of Hepatitis C in symptomatic patients.
• It is part of routine screening in blood donors and pre-surgical evaluations.
✓. Consultation:
Patients with a positive result are to be advised to consult a healthcare provider for confirmatory testing (e.g., HCV RNA PCR) to assess active infection. Counseling on liver health, transmission prevention, and antiviral treatment options was recommended.

24/07/2025

1. Malaria Test

1. Objective:

The objective of the malaria test is to detect the presence of Plasmodium species in the blood of suspected individuals to confirm malaria infection.

2. Principle:

The test is based on detecting malaria parasites in blood either by microscopic examination of stained blood smears or through rapid diagnostic tests (RDTs) which identified specific antigens of Plasmodium species.

3. Materials:

Capillary or venous blood

Sterile lancet

Glass slides (for microscopy)

Giemsa stain

Microscope

RDT kit (cassette, buffer)

Gloves and PPE

Timer and waste disposal materials

4. Procedure (Microscopic Method):

1. A thick and thin blood smear was prepared on a slide using a fresh blood sample.

2. The smear was air-dried and stained with Giemsa stain.

3. The slide was examined under a microscope using oil immersion.

4. Parasites were identified by morphology and counted for parasite load.

4. Procedure (RDT):

1. A finger-prick blood sample was applied to the RDT cassette.

2. Buffer was added and the cassette was left to develop.

3. Test results were read within 15–20 minutes based on line visibility (control and test lines).

5. Result:

Positive: Parasites or antigens of Plasmodium spp. detected.

Negative: No parasites or antigens detected.
Species may include P. falciparum, P. vivax, etc.

6. Uses:

Confirm diagnosis of malaria

Guide immediate treatment

Monitor treatment success

Support public health surveillance

2. Implication to the Community:

Positive Impacts:

Early detection reduces severe malaria cases and deaths.

Prevents outbreaks by identifying and isolating sources.

Guides vector control (e.g., insecticide spraying, net distribution).

Informs health policies for resource allocation and education.

Encourages health-seeking behavior through awareness.

Risks of Poor Testing:

Misdiagnosis can lead to untreated malaria or drug misuse.

Inaccurate results increase community spread and resistance.

Delay in detection burdens healthcare systems.

24/07/2025

Hepatitis B Profile Test

1. Objective:

The objective of the Hepatitis B profile test is to detect and evaluate infection status, immune response, and chronicity of Hepatitis B virus (HBV) in a patient.

2. Principle:

The test is based on ELISA (enzyme-linked immunosorbent assay) or rapid immunochromatographic methods, which detected HBV antigens and antibodies using specific binding and visual color development.

3. Materials:

Patient’s blood sample (serum or plasma)

HBV profile test kit (ELISA or rapid strip)

Micropipette and tips

Test tubes and reagents

Incubator (for ELISA)

ELISA reader (if applicable)

Gloves and protective gear

4. Proceduer

1. Blood is collected aseptically and serum is separated.

2. Test strips or ELISA wells are prepared with required reagents.

3. Serum is added to each test zone (e.g., HBsAg, HBeAg, anti-HBs, anti-HBe, anti-HBc).

4. The test was incubated (e.g., 15–30 mins for rapid tests or 1–2 hours for ELISA).

5. Color development or band formation observed.

6. The presence or absence of each marker are recorded.

5. Result:

Marker Meaning

HBsAg (+) Active infection (acute or chronic)
HBeAg (+) High infectivity
Anti-HBc (+) Previous or ongoing infection
Anti-HBs (+) Immunity (past infection or vaccination)
Anti-HBe (+) Recovery or low infectivity phase

6. Uses:

Diagnose acute or chronic Hepatitis B infection

Monitor disease stage and infectivity

Screen for HBV in blood donors

Determine immunity post-vaccination

Guide treatment and monitoring decisions

7. Consultation:

Patients with positive HBsAg are to be advised to undergo further tests such as HBV DNA PCR, liver function tests (LFTs), and liver ultrasound. Proper medical guidance, antiviral therapy (if needed), and lifestyle changes are recommended.

21/07/2025

Urinalysis

👉What is Urinalysis?
Urinalysis is a common test that analyzes urine to detect and manage various health conditions. It checks for abnormalities, signs of infection, or other indicators of disease.

👉Components of Urinalysis
1. Physical Examination: Looks at urine color, clarity, and concentration (specific gravity).
2. Chemical Examination: Uses dipsticks to test for substances like:
- pH: Urine acidity.
- Protein: High levels may indicate kidney damage.
- Glucose: Presence could suggest diabetes.
- Ketones: May indicate diabetes or fasting.
- Blood: Could signal infection, stones, or injury.
3. Microscopic Examination: Checks for cells, crystals, bacteria, or other elements like:
- Red blood cells: Possible kidney or urinary tract issues.
- White blood cells: May indicate infection.
- Bacteria: Suggests urinary tract infection.

👉 Uses of Urinalysis
- Diagnose urinary tract infections (UTIs): Detects bacteria or white blood cells.
- Check kidney function: Looks for protein or blood.
- Monitor diabetes: Checks for glucose or ketones.
- Detect substance use: Some tests check for drugs.

15/07/2025

Erythrocyte Sedimentation Rate (ESR) Test
1. Objective
The objective of the ESR test was to measure the rate at which red blood cells (RBCs) settled at the bottom of a vertical tube in a given time period (usually 1 hour), to assess the presence of inflammation or infection in the body.

2. Principle
The ESR test was based on the principle that inflammatory processes caused red blood cells to clump together (rouleaux formation), increasing their sedimentation rate.
• The greater the inflammation, the faster the RBCs settled.
• The distance (in mm) that RBCs fell in a vertical tube over 1 hour indicated the ESR value.

3. Materials
• Fresh anticoagulated blood (with EDTA or sodium citrate)
• Westergren ESR tube (200 mm long)
• Westergren pipette stand or ESR rack
• Timer or stopwatch
• Gloves
• Syringe or pipette

4. Procedure (Macroscopic)
1. 1 part of 3.8% sodium citrate was mixed with 4 parts of blood.
2. The mixture was drawn into a Westergren tube up to the 0 mark.
3. The tube was placed vertically in an ESR rack.
4. After exactly 1 hour, the level of plasma above the RBC column was recorded in millimeters.

5. Result
• ESR was recorded in mm/hr.
• Normal Range:
o Males: 0–15 mm/hr
o Females: 0–20 mm/hr
o Children: 0–10 mm/hr
o Elderly: Slightly higher
• Elevated ESR indicated inflammation, infection, autoimmune disorders, or malignancy.

6. Uses
• Used as a non-specific marker of inflammation.
• Helped monitor disease progression or treatment effectiveness in conditions like:
o Rheumatoid arthritis
o Tuberculosis
o Temporal arteritis
o Chronic infections
•Also used in diagnosing autoimmune diseases and certain cancers.

7. Consultation
• ESR results were interpreted alongside clinical symptoms and other laboratory findings.
• A high ESR alone did not confirm a diagnosis but prompted further investigations.
• Physicians used ESR to monitor chronic conditions and determine disease activity.

09/07/2025

Widal Test

1. Objective

The objective of the Widal test is to detect agglutinating antibodies (agglutinins) against the O and H antigens of Salmonella typhi and Salmonella paratyphi, aiding in the diagnosis of enteric (typhoid and paratyphoid) fever.

2. Principle

The Widal test is a serological agglutination test. When serum from a typhoid patient is mixed with killed Salmonella antigens, it caused visible clumping (agglutination) if specific antibodies were present.

3. Materials

Patient’s serum sample

Widal antigen suspensions (O, H, AH, BH)

Test tubes or slide for slide agglutination

Pipettes

Saline

Water bath (optional for incubation)

Slide/Tube agglutination viewer

4. Procedure

A. Slide Method (Qualitative)

1. A drop of patient’s serum is placed on two(O,H) or four(O,H,AH,BH) cycle Mark slide

2. Equal volumes of each antigen (O, H, AH, BH) are added.

3. The slide is rotated for 1 minute.

4. And Agglutination (clumping) is observed.

B. Tube Method (Quantitative) Dilution method

1. Serum is serially diluted in Five test tubes.

2. Antigen suspensions are added to each tube according to the reaction ratio as 1:20,1:40,1:80,1:160 and 1:320.

3. Tubes are incubated at 37°C for 16–20 hours.

4. The highest dilution showing agglutination is recorded as the antibody titer.

5. Result

Significant titers:

Anti-O: ≥1:320

Anti-H: ≥1:320

A rising titer in paired samples over 7–10 days is more diagnostic.

Note: A single high titer may not confirm active infection without clinical correlation or history of vaccination.

6. Uses

Diagnosed typhoid and paratyphoid fever

Monitored antibody response

Common in areas with limited access to culture facilities

7. Conclusion

The Widal test provided a quick and simple serological method for presumptive diagnosis of enteric fever, though it required careful interpretation and correlation with clinical findings and other tests.

09/07/2025

Acid-Fast Staining (AFB Stain)
1. Objective
To detect acid-fast bacilli (AFB), especially Mycobacterium species, in clinical specimens such as sputum, stool, or tissue, using the Ziehl-Neelsen or Kinyoun staining method.

2. Principle
Acid-fast bacteria (AFB) possess mycolic acid in their cell walls, making them resistant to decolorization by acid-alcohol. In Ziehl-Neelsen staining, AFB retain the primary stain (carbol fuchsin) after acid-alcohol treatment and appear bright red against a blue background.

3. Materials
• Clean glass microscope slides
• Carbol fuchsin stain
• Acid-alcohol (decolorizer: 3% HCl in ethanol)
• Methylene blue or malachite green (counterstain)
• Bunsen burner or slide warmer (for heat-fixing)
• Staining rack and forceps
• Microscope (preferably oil immersion lens)
• PPE: gloves, mask, lab coat

4. Procedure (Ziehl-Neelsen Method)
1. Prepare a smear of the clinical specimen on a clean slide.
2. Air dry and heat fix the slide.
3. Flood the slide with carbol fuchsin and heat gently until steaming (do not boil) for 5 minutes.
4. Rinse with water.
5. Decolorize with acid-alcohol for 2–3 minutes or until the slide runs clear.
6. Rinse again with water.
7. Counterstain with methylene blue (or malachite green) for 1–2 minutes.
8. Rinse, air dry, and examine under oil immersion (100x).
Kinyoun method is a cold stain (no heating) alternative using a higher concentration of carbol fuchsin.

5. Result Interpretation
Observation Interpretation
Bright red rods (AFB) Positive for acid-fast bacilli (e.g., Mycobacterium tuberculosis)
Blue background only Negative for AFB

6. Uses
• Diagnosis of tuberculosis (pulmonary and extrapulmonary)
• Identification of nontuberculous mycobacteria (NTM)
• Detection of Cryptosporidium or Isospora in stool (modified acid-fast)
• Public health surveillance and TB control

7. Conclusion
Acid-fast staining is a rapid and specific method for detecting mycobacteria and other acid-fast organisms. While less sensitive than molecular tests, it remains critical for initial TB screening, especially in resource-limited settings.

08/07/2025

Gram Staining test

1. Objective
The objective of the test was to differentiate bacterial species into Gram-positive and Gram-negative groups based on the chemical and physical properties of their cell walls, aiding in identification and treatment decisions.

2. Principle
Gram staining was based on the differential retention of crystal violet dye.

Gram-positive bacteria retained the crystal violet-iodine complex and appeared purple due to their thick peptidoglycan layer.

Gram-negative bacteria, with a thin peptidoglycan layer and outer membrane, lost the violet stain and were counterstained pink/red with safranin.

3. Materials
Bacterial smear on glass slide

Crystal violet (primary stain)

Gram’s iodine (mordant)

Alcohol or acetone (decolorizer)

Safranin (counterstain)

Microscope

Staining rack and water supply

4. Procedure
A thin smear of the bacterial sample was heat-fixed on a glass slide.

The slide was flooded with crystal violet for 1 minute, then rinsed.

Gram's iodine was applied for 1 minute to form a complex with the dye.

The slide was briefly decolorized with alcohol/acetone, then rinsed.

It was counterstained with safranin for 30–60 seconds.

After final rinsing, the slide was dried and observed under oil immersion microscopy.

5. Result
Gram-positive bacteria: Appeared purple (e.g., Staphylococcus aureus, Streptococcus spp.)

Gram-negative bacteria: Appeared pink/red (e.g., E. coli, Pseudomonas aeruginosa)

Mixed or no staining could suggest contamination or faulty technique.

6. Uses
Rapid preliminary bacterial classification

Guided antibiotic therapy (Gram-negative often more resistant)

Differentiated between types of infection

Used in clinical, environmental, and food microbiology

7. Conclusion
Gram staining was a fundamental microbiological technique that allowed quick and valuable differentiation between Gram-positive and Gram-negative bacteria, facilitating effective diagnosis and treatment decisions.

08/07/2025

H. pylori Antigen in Stool test
1. Objective
The objective of this test was to detect the presence of Helicobacter pylori (H. pylori) antigen in a stool sample, which indicated an active infection in the stomach lining.

2. Principle
The test used immunochromatographic assay or enzyme immunoassay (EIA/ELISA) to identify H. pylori-specific antigens in stool. A positive reaction occurred when the antigen in the stool bound to monoclonal antibodies immobilized on a test strip or microwell, producing a visible color change or line.

3. Materials
• Fresh stool sample in a sterile container
• H. pylori antigen test kit (rapid or ELISA)
• Sample dilution buffer
• Dropper or micropipette
• Rapid test cassette or ELISA plate
• Gloves, PPE, and timer
• ELISA reader (if using EIA method)

4. Procedure (Rapid Test Format)
1. A small quantity of stool was mixed with the provided buffer.
2. A few drops of the prepared sample were added to the sample well of the test cassette.
3. The result was read after 10–15 minutes:
o Two lines = Positive
o One line (control only) = Negative
o No lines or one test line only = Invalid
4. In ELISA, sample and reagents were added in sequence, and optical density (OD) was read at 450 nm.

5. Result (Example)
Test Type Result
H. pylori Antigen Positive (two lines)
🧪 Interpretation: The test confirmed an active H. pylori infection, commonly associated with gastritis, peptic ulcer, or duodenal ulcer.

6. Uses
• Diagnosed active H. pylori infection
• Helped monitor treatment success or failure
• Used in evaluating patients with upper abdominal symptoms
• Preferred non-invasive method, especially in children

7. Conclusion
The H. pylori antigen in stool test was a highly specific, rapid, and non-invasive method for detecting active H. pylori infection. It played a key role in the diagnosis and follow-up of patients with dyspepsia and peptic ulcers.

18/02/2025