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KenMartin E-Pro Diagnostic, chidoziekenmartin@gmail. com, Enugu (2025)

17/10/2025

DNA Test

1. Objective:

The test was performed to identify and analyze genetic material (DNA) for purposes such as paternity testing, genetic disease detection, or forensic identification.

2. Principle:

The test was based on the extraction of DNA from cells, followed by amplification and analysis of specific DNA regions using techniques such as Polymerase Chain Reaction (PCR) or electrophoresis. Each person’s DNA pattern was unique, allowing identification and comparison.

3. Materials:

• Blood, saliva, or buccal swab sample
• DNA extraction kit or reagents (lysis buffer, ethanol, etc.)
• Microcentrifuge tubes
• Micropipettes and tips
• PCR reagents (primers, Taq polymerase, nucleotides, buffer)
• Agarose gel and electrophoresis apparatus
• UV transilluminator or gel documentation system.

4. Procedure (Microscopic/Analytical):

1. The biological sample was collected and labeled properly.
2. DNA was extracted using chemical or enzymatic methods.
3. The purity and concentration of DNA were checked using a spectrophotometer.
4. Specific DNA segments were amplified by PCR.
5. The amplified DNA was separated on an agarose gel by electrophoresis.
6. The banding pattern was visualized under UV light and compared with control or reference samples.

5. Result:

Clear DNA bands were observed on the gel under UV light. The presence, absence, or pattern of bands indicated genetic similarity or differences, depending on the test purpose (e.g., match in paternity testing or mutation detection).

6. Uses:

• To establish paternity or family relationships.
• To identify genetic mutations or inherited diseases.
• To assist in forensic investigations.
• To study genetic diversity and research applications.

7. Consultation:

Results were interpreted by a qualified geneticist or laboratory specialist. Individuals were advised to discuss findings with a medical genetic counselor or physician for accurate understanding and further advice.

13/10/2025

Semen Analysis (Semen Test)
1. Objective:
The objective of the semen test was to evaluate the quantity and quality of a man’s semen and s***m to assess fertility, confirm successful vasectomy, or investigate reproductive or hormonal disorders.
2. Principle:
The test was based on microscopic and physical examination of semen to assess s***m count, motility, morphology, and viability. It provided information about the functional status of the male reproductive system, including the te**es, seminal vesicles, and prostate gland.
3. Materials:
• Sterile, wide-mouthed collection container
• Clean glass slides and cover slips
• Microscope
• Graduated pipette or semen analyzer
• Timer
• Stains (e.g., Eosin–Nigrosin for morphology)
• pH paper
• Water bath (37°C)
4. Procedure (Microscopic and Physical Examination):
1. The semen sample was collected after 2–5 days of abstinence, by ma********on into a sterile container.
2. The sample was allowed to liquefy at room temperature (within 30 minutes).
3. Physical examination:
o Volume, color, viscosity, and pH were recorded.
4. Microscopic examination:
o A drop of semen was placed on a slide and observed under a microscope.
o S***m count was determined using a counting chamber (e.g., Neubauer).
o S***m motility was assessed by observing active movement.
o S***m morphology was examined after staining.
o Presence of pus cells or abnormal cells was noted.
5. Result:
• Normal Semen Parameters (WHO reference values):
o Volume: 1.5–6.0 mL
o pH: 7.2–8.0
o S***m count: ≥15 million/mL
o Motility: ≥40% (progressive + non-progressive)
o Morphology: ≥4% normal forms
o Liquefaction: within 30 minutes
6. Uses:
• To assess male fertility potential
• To confirm effectiveness of vasectomy
• To diagnose causes of infertility (e.g., oligos***mia, asthenos***mia, azoos***mia)
• To evaluate testicular or accessory gland function
7. Consultation:
Patients with abnormal semen results were advised to consult a urologist or fertility specialist. Further investigations such as hormone assays, scrotal ultrasound, or genetic testing could be recommended to identify the underlying cause and guide appropriate treatment.

08/10/2025

SS Agar Test

Objective:

To isolate and differentiate Salmonella and Shigella species from other enteric bacteria using SS agar medium.

Principle:

SS agar is both selective and differential.

Bile salts and brilliant green dye inhibit most Gram-positive and many Gram-negative bacteria.

Lactose and the pH indicator neutral red differentiate lactose fermenters (pink colonies) from non-fermenters (colorless colonies).

Ferric citrate and sodium thiosulfate detect hydrogen sulfide (H₂S) production (black-centered colonies).

Materials Required:

SS agar plates

Sterile inoculating loop

Bacterial cultures or stool sample

Incubator (37°C)

Sterile saline or peptone water (for dilution)

Procedure:

1. Label SS agar plates with sample details.

2. Streak the plate using the quadrant streak method for isolation.

3. Incubate at 37°C for 24–48 hours.

4. Observe colony morphology.

Observation:

Salmonella: Colorless colonies with black centers (H₂S production).

Shigella: Colorless colonies without black centers.

Lactose fermenters (e.g., E. coli): Pink to red colonies.

Result/Conclusion:

The presence of non-lactose fermenting colonies (especially with or without H₂S) suggests possible Salmonella or Shigella species.

Further biochemical or serological confirmation is required.

Precautions:

Avoid contamination during streaking.

Do not over-incubate (may alter colony color).

Use fresh culture media.

06/10/2025

Semen Analysis Test
1. Objective:
The objective of this test was to evaluate the quality and quantity of semen to assess male fertility potential. It determined s***m count, motility, morphology, and overall seminal fluid characteristics.
2. Principle:
The test was based on the microscopic examination of semen to measure parameters such as s***m concentration, motility, and morphology. The evaluation followed WHO guidelines, where the semen sample was observed under the microscope to determine s***m health and fertility status.
3. Materials:
• Fresh semen sample (collected in sterile container)
• Microscope and slides
• Coverslips
• Graduated pipette
• Neubauer counting chamber (hemocytometer)
• Staining reagents (Eosin-Nigrosin or Papanicolaou stain)
• Gloves and disinfectant
4. Procedure (Microscopic):
1. The semen sample was collected after 3–5 days of abstinence and allowed to liquefy for 30 minutes at room temperature.
2. The physical characteristics (volume, color, viscosity, and pH) were noted.
3. A drop of semen was placed on a clean slide, covered with a coverslip, and examined under low and high power for motility and morphology.
4. For s***m count, the sample was diluted and loaded into a Neubauer counting chamber, and s***m cells were counted under the microscope.
5. A stained smear was prepared for morphological assessment.
5. Result:
• Normal: Volume 2–6 mL, s***m count >15 million/mL, motility >40%, normal morphology >4%, and pH 7.2–8.0.
• Abnormal: Low s***m count (oligos***mia), no s***m (azoos***mia), low motility (asthenozoos***mia), or abnormal morphology (teratozoos***mia).
6. Uses:
• It was used to evaluate male infertility.
• It helped monitor s***m recovery after treatment.
• It aided in assessing the effects of hormonal or testicular disorders.
7. Consultation:
Men with abnormal semen parameters were advised to consult a urologist or fertility specialist. Further investigations like hormonal analysis, ultrasound, or genetic tests were recommended depending on findings.

30/09/2025

Coombs Test (Direct and Indirect Antiglobulin Test)

1. Objective:
The objective of the Coombs test was to detect the presence of antibodies or complement proteins bound to the surface of red blood cells (RBCs). The Direct Coombs Test (DAT) aimed to identify antibodies attached directly to the patient's RBCs, while the Indirect Coombs Test (IAT) was performed to detect free antibodies present in the patient's serum that could react with RBC antigens.

2. Principle:
The principle of the Coombs test was based on the detection of antigen–antibody reactions. In the Direct Coombs Test, RBCs coated with immunoglobulin (IgG) or complement were mixed with Coombs reagent (anti-human globulin). The reagent cross-linked the sensitized RBCs, causing visible agglutination.
In the Indirect Coombs Test, the patient's serum containing antibodies was first incubated with donor or test RBCs. If the antibodies in the serum bound to the antigens on the RBCs, the subsequent addition of Coombs reagent caused agglutination. The appearance of agglutination indicated a positive test result.

3. Materials:
• Patient's blood sample (for Direct Coombs Test)
• Patient's serum (for Indirect Coombs Test)
• Test RBCs (antigen-positive cells for IAT)
• Coombs reagent (anti-human globulin serum)
• Test tubes
• Pipettes
• Centrifuge
• Microscope
• Isotonic saline solution
• Glass slides

4. Procedure (Microscopic):
a) Direct Coombs Test (DAT):
1. A blood sample was collected from the patient, and RBCs were washed three times with isotonic saline to remove unbound antibodies.
2. The washed RBC pellet was suspended in saline to make a 2–5% RBC suspension.
3. Two drops of Coombs reagent were added to the washed RBC suspension in a test tube.
4. The tube was mixed gently and centrifuged for 1–2 minutes at 1000 rpm.
5. The sediment was examined microscopically for agglutination.
6. The presence of clumping indicated a positive Direct Coombs Test.
b) Indirect Coombs Test (IAT):
1. Two drops of the patient’s serum were mixed with one drop of test RBC suspension in a test tube.
2. The mixture was incubated at 37°C for 30 minutes to allow antibodies to bind to RBC antigens.
3. After incubation, the RBCs were washed three times with saline to remove unbound antibodies.
4. Two drops of Coombs reagent were added to the washed RBCs.
5. The tube was centrifuged for 1–2 minutes and then examined microscopically.
6. Agglutination of RBCs indicated a positive Indirect Coombs Test.

5. Result:
• Positive Result: Visible clumping or agglutination of RBCs was observed under the microscope, indicating the presence of antibodies bound to RBCs (DAT) or free antibodies in the serum (IAT).
• Negative Result: No agglutination was observed, indicating the absence of significant antibodies.

6. Uses:
• To diagnose autoimmune hemolytic anemia by detecting autoantibodies attached to the patient's RBCs.
• To screen for hemolytic disease of the newborn (HDN) caused by Rh incompatibility between mother and fetus.
• To cross-match blood during transfusion to prevent hemolytic transfusion reactions.
• To detect antibodies in patients receiving multiple transfusions.
• To monitor patients for immune-mediated drug reactions affecting RBCs.

7. Consultation:
If the test result was positive, the patient was advised to consult a hematologist or immunologist for further evaluation and management. In cases of suspected hemolytic disease of the newborn, immediate neonatal care was recommended. For blood transfusion-related incompatibilities, the blood bank physician was consulted to ensure safe transfusion practices. The patient was counseled about possible causes and preventive measures, including Rh immunoglobulin administration in Rh-negative mothers.

27/09/2025

MacConkey Agar Test

1. Objective:
To isolate and differentiate Gram-negative enteric bacteria based on their ability to ferment lactose.

2. Principle:

MacConkey agar is both selective and differential.

Selective agent: Bile salts and crystal violet inhibit Gram-positive bacteria.

Differential agent: Lactose and neutral red indicator.

Lactose fermenters → produce acid → colonies turn pink/red.

Non-lactose fermenters → no acid → colonies remain colorless or pale.

3. Materials:

MacConkey agar plates

Inoculating loop

Bunsen burner/spirit lamp

Test bacterial culture

Incubator (35–37 °C)

4. Procedure:

1. Sterilize the inoculating loop.

2. Pick a small amount of bacterial culture.

3. Streak the culture on the surface of a MacConkey agar plate.

4. Incubate at 35–37 °C for 18–24 hours.

5. Observe colony growth and color change.

5. Result Interpretation:

Pink colonies: Lactose fermenters (e.g., E. coli).

Colorless/transparent colonies: Non-lactose fermenters (e.g., Salmonella, Shigella).

No growth: Gram-positive bacteria are inhibited.

6. Uses:

Differentiating enteric bacteria (lactose vs non-lactose fermenters).

Identifying enteropathogens like Salmonella and Shigella.

Routine diagnostic bacteriology in clinical and food microbiology.

7. Conclusion:
MacConkey agar is a simple, reliable medium for the isolation and differentiation of Gram-negative enteric bacteria based on lactose fermentation.

15/09/2025

Forward and Reverse Blood Grouping System

1. Objective
• The objective of the blood grouping test was to determine an individual’s ABO and Rh blood group using both forward (cell grouping) and reverse (serum grouping) methods.
• It was essential for safe blood transfusion, organ transplantation, and pregnancy management.

2. Principle
The test was based on the principle of antigen–antibody reaction.
• Forward grouping (cell grouping): Patient’s red cells were tested with known antisera (Anti-A, Anti-B, Anti-D). Agglutination indicated the presence of corresponding antigen.

• Reverse grouping (serum grouping): Patient’s serum was tested with known reagent red cells (A cells, B cells). Agglutination indicated the presence of corresponding antibody.

• Forward and reverse results complemented each other for confirmation.

3. Materials
1. Patient’s blood sample (EDTA tube for cells, plain tube for serum)
2. Reagent antisera: Anti-A, Anti-B, Anti-D
3. Known red cells: A cells, B cells
4. Glass slides or microplate
5. Test tubes and racks
6. Pasteur pipettes/micropipettes
7. Centrifuge
8. Microscope (to confirm weak agglutination)

4. Procedure
Forward Grouping (Cell Grouping):
• A drop of Anti-A, Anti-B, and Anti-D antisera was placed separately on a slide.
• A drop of patient’s red cell suspension was added to each.
• The mixture was gently rocked and observed for agglutination.
Reverse Grouping (Serum Grouping):
• Patient’s serum was mixed with known A cells and B cells in separate tubes.
• After centrifugation, the mixture was observed for agglutination.
• Results were cross-checked with forward grouping.
5. Result
• Example (Forward Grouping):
o Agglutination with Anti-A → A antigen present.
o Agglutination with Anti-B → B antigen present.
o Agglutination with Anti-D → Rh-positive.
• Example (Reverse Grouping):
o Agglutination with B cells → Anti-B antibody present → Group A.
o Agglutination with A cells → Anti-A antibody present → Group B.
• Concordance between forward and reverse grouping confirmed the blood type (e.g., A+, B–, O+, AB+).

6. Uses
• It was used to determine safe donor–recipient compatibility in transfusion.
• It was useful in antenatal screening for hemolytic disease of the newborn.
• It helped in forensic investigations and medico-legal cases.
• It was applied in organ and tissue transplantation.

7. Conclusion
• The combination of forward and reverse grouping gave an accurate determination of a person’s blood group.
• This dual system minimized errors and ensured safety in transfusion and clinical practices.

06/09/2025

Sickle Cell Test (Sickling Test / Solubility Test)
1. Objective
The objective of the sickle cell test (also known as the sickling test or solubility test) was to screen for sickle cell disease or the sickle cell trait by detecting the presence of sickle-shaped red blood cells under specific conditions.
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2. Principle
The test was based on the principle that hemoglobin S (HbS), found in individuals with sickle cell disease or sickle cell trait, forms insoluble polymers under low oxygen conditions. These polymers distort the red blood cells, causing them to assume a sickle shape, which can be visualized under a microscope. The solubility test uses a reagent to lower oxygen tension and observe the precipitation of hemoglobin S.
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3. Materials
• Fresh blood sample (preferably venous)
• Sodium metabisulfite or phosphate buffer solution
• Microscope and glass slides
• Test tubes and pipettes
• Solubility reagent (sodium metabisulfite solution or similar)
• Control samples (normal hemoglobin and sickle cell disease blood samples)
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4. Procedure
1. A blood sample was collected from the patient.
2. A small amount of blood was mixed with a sickling reagent (sodium metabisulfite solution) in a test tube.
3. The mixture was incubated at room temperature for 10-15 minutes.
4. After incubation, the sample was examined under a microscope for the presence of sickle-shaped red blood cells.
5. For the solubility test: A few drops of the sample were added to a special reagent in a test tube. If HbS was present, the solution would appear turbid or cloudy due to the precipitation of hemoglobin S.
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5. Results
• Positive result (Sickle Cell Disease or Trait):
o Sickle-shaped red blood cells are visible under the microscope, or the solution becomes turbid, indicating the presence of hemoglobin S.
• Negative result:
o No sickle-shaped red blood cells or turbidity observed in the solution.
________________________________________
6. Uses
• Screening for sickle cell disease and sickle cell trait.
• To diagnose hemoglobinopathies in individuals with suspected sickle cell disease or trait.
• To confirm the presence of HbS in newborns or individuals with a family history of sickle cell disease.
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7. Consultation
• A positive result should be confirmed with hemoglobin electrophoresis or HPLC for definitive diagnosis.
• Genetic counseling and family planning advice are often recommended for individuals with sickle cell trait.
• Patients with sickle cell disease require ongoing medical management to prevent complications such as pain crises, infections, and organ damage.

05/09/2025

High Vaginal Swab (HVS) Test
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1. Objective

The objective of the HVS test was to detect infections in the female ge***al tract by isolating and identifying bacteria, fungi, or parasites from a va**nal swab.

2. Principle

The test was based on the principle that microorganisms causing va**nal infections (such as Candida albicans, Trichomonas va**nalis, Neisseria gonorrhoeae, or bacterial vaginosis organisms) could be detected by direct microscopy, culture, and biochemical tests.

3. Materials

Sterile va**nal swab stick

Transport medium (Amies or Stuart medium)

Glass slides and cover slips

Microscope

Gram stain and saline preparation reagents

Culture media (Blood agar, MacConkey agar, Sabouraud agar, Chocolate agar)

Incubator (35–37°C)

4. Procedure

Sample Collection:

1. A sterile swab was inserted into the posterior fornix of the va**na.

2. The swab was placed into transport medium and sent to the lab.

Microscopic Examination:

1. A smear was prepared from the swab.

2. Saline wet mount was observed for motile Trichomonas va**nalis.

3. KOH preparation was examined for fungal elements (Candida).

4. Gram stain was performed to check bacterial morphology and clue cells (for bacterial vaginosis).

Culture:

1. The swab was streaked onto Blood agar, MacConkey agar, and Sabouraud agar.

2. Plates were incubated at 35–37°C.

3. Growth was identified based on colony morphology, Gram stain, and biochemical tests.

5. Results

Normal (Negative): Predominantly lactobacilli with no pathogenic organisms.

Abnormal (Positive):

Yeast cells/pseudohyphae → Candida albicans

Motile flagellated organisms → Trichomonas va**nalis

Gram-negative diplococci → Neisseria gonorrhoeae

Clue cells → Bacterial vaginosis

6. Uses

To diagnose causes of va**nal discharge, itching, or odor.

To detect sexually transmitted infections (STIs).

To differentiate between bacterial, fungal, and parasitic infections.

To guide appropriate treatment.

7. Consultation

Positive results required consultation with a gynecologist for specific antimicrobial or antifungal therapy.

Sexual partners were also advised treatment in case of STIs.

Negative results but persistent symptoms required further testing, such as PCR for Chlamydia trachomatis or Mycoplasma.

23/08/2025

RBC Count (Red Blood Cell Count) Test

1. Objective:
The objective of the RBC Count test was to determine the number of red blood cells (RBCs) in a blood sample. This test is crucial for diagnosing conditions such as anemia, polycythemia, and other blood disorders that affect the red blood cell population.

2. Principle:
The RBC Count test is typically performed using an automated hematology analyzer or manually with a microscope. The principle is based on counting the RBCs in a given volume of blood. In automated methods, the analyzer uses light scattering or electrical impedance to count and size the cells. In manual methods, a hemocytometer is used, and the RBCs are counted under a microscope after dilution with a special solution.

3. Materials:
o Blood sample (usually collected in an EDTA tube)
o Automated hematology analyzer or hemocytometer
o Dilution fluid (e.g., saline or a special RBC counting solution)
o Microscope (for manual methods)
o Pipettes and test tubes

4. Procedure:
o Automated Method:
 A small volume of the blood sample is introduced into an automated hematology analyzer.
 The analyzer measures the number of RBCs by either light scattering or electrical impedance techniques.
 The results are displayed directly on the analyzer's screen.
o Manual Method (using a hemocytometer):
 A drop of blood is diluted with a special solution that lyses white blood cells and preserves red blood cells.
 A known volume of the diluted blood is placed on a hemocytometer.
 The RBCs are counted in a specific grid pattern under a microscope, and the concentration is calculated based on the dilution and the number of cells counted.

5. Result:
The RBC count result is expressed as the number of red blood cells per microliter of blood (cells/μL) or per liter (cells/L). Normal RBC counts generally range from:
o Men: 4.7–6.1 million cells/μL
o Women: 4.2–5.4 million cells/μL
o Children: 4.1–5.5 million cells/μL
Abnormal results can indicate conditions like anemia (low RBC count), polycythemia (high RBC count), or other hematological disorders.

6. Uses:
o The RBC Count is primarily used to evaluate the health of the red blood cells and to diagnose conditions such as:
 Anemia (low RBC count)
 Polycythemia (high RBC count)
 Hemorrhage (due to blood loss)
 Chronic obstructive pulmonary disease (COPD)
 Dehydration (can cause falsely elevated RBC count)
o It is also part of a routine complete blood count (CBC) to evaluate overall health.

7. Consultation:
Abnormal RBC counts are typically followed by consultation with a hematologist or a healthcare provider. Further diagnostic tests, such as hemoglobin and hematocrit measurements, iron studies, or bone marrow biopsy, may be recommended to confirm the cause of the abnormal RBC count and guide treatment. Depending on the diagnosis, treatment options could include iron supplementation, blood transfusion, or medications to address the underlying condition.

09/08/2025

OGTT (Oral Glucose Tolerance Test) is a laboratory test used to check how well your body handles glucose (sugar) and to help diagnose conditions like diabetes, gestational diabetes, and prediabetes.
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Purpose

Diagnose type 2 diabetes.

Detect gestational diabetes in pregnancy.

Identify prediabetes.

Evaluate glucose metabolism disorders.
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Principle

The OGTT measures how quickly glucose is cleared from the blood after drinking a measured glucose solution. Normally, blood glucose rises after ingestion, but returns to near normal within 2 hours as insulin facilitates glucose uptake.
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Procedure

Preparation (Before the Test):

Fast for at least 8–12 hours (only water allowed).

Avoid smoking, alcohol, and excessive exercise before the test.

Eat a balanced diet (≥150 g carbohydrate/day) for 3 days before the test.

Steps:

1. Fasting Blood Sample — Blood is drawn after overnight fasting to measure baseline glucose.

2. Glucose Drink — Patient drinks a solution containing 75 g anhydrous glucose (for adults) dissolved in 250–300 mL of water, consumed within 5 minutes.

For pregnant women: 75 g glucose (WHO recommendation) or 100 g (some protocols).

For children: 1.75 g/kg body weight (max 75 g).

3. Blood Sampling — Blood glucose levels are measured at specific intervals:

0 min (fasting)

30 min (optional)

1 hour

2 hours

4. Interpretation — Compare results with reference values.
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Interpretation (WHO Criteria for 75 g OGTT in non-pregnant adults)

Time (hours) Normal Impaired Glucose Tolerance (Prediabetes) Diabetes

Fasting < 100 mg/dL (< 5.6 mmol/L) 100–125 mg/dL (5.6–6.9 mmol/L) ≥ 126 mg/dL (≥ 7.0 mmol/L)
2 hours < 140 mg/dL (< 7.8 mmol/L) 140–199 mg/dL (7.8–11.0 mmol/L) ≥ 200 mg/dL (≥ 11.1 mmol/L)
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For Gestational Diabetes (75 g OGTT, WHO 2013)

Only one abnormal value is enough for diagnosis:

Time Threshold

Fasting ≥ 92 mg/dL (≥ 5.1 mmol/L)
1 hour ≥ 180 mg/dL (≥ 10.0 mmol/L)
2 hours ≥ 153 mg/dL (≥ 8.5 mmol/L)
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Advantages

Gold standard for diagnosing gestational diabetes and glucose intolerance.

Detects abnormalities earlier than fasting glucose alone.

Limitations

Time-consuming (2–3 hours).

Requires patient preparation and compliance.

Can cause nausea or dizziness after drinking glucose solution.

07/08/2025

Precision begins at the tip. Pipetting is the heartbeat of accurate diagnostics in the medical laboratory. With every measured drop, it ensures reliability in test results, from blood chemistry to molecular assays.

KenMartin E-Pro Diagnostic

17/10/2025

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