27/10/2025
G6PD deficiency testing
Bottom line
G6PD enzyme assays detect decreased red-cell G6PD activity. Timing and method matter: testing during acute hemolysis or after transfusion can produce false-normal results. Know when to run the assay, which method to use, and when to request molecular testing.
When to order
• Suspected acute hemolysis after oxidant exposure (drugs, fava beans, infections).
• Neonatal jaundice with hemolysis features.
• Family history of G6PD deficiency or recurrent episodic hemolysis.
Timing & specimen rules
• Avoid testing during acute hemolysis if possible. Reticulocytosis and young RBCs have higher G6PD activity and can mask deficiency. Wait until the reticulocyte count normalizes (often ≥2–3 months) when feasible.
• If an urgent result is needed during hemolysis, perform testing but flag the result as potentially false-normal and recommend repeat testing after recovery or perform molecular testing.
• Recent transfusion dilutes patient RBCs and can give false-normal results; avoid testing until transfused RBCs are cleared, or use genetic testing.
• Collect EDTA whole blood, run the assay promptly (same day) or follow manufacturer instructions for sample stability. Document collection time and any recent transfusion/hemolysis on the requisition.
Available tests & their uses
• Qualitative fluorescent spot test — fast screening; sensitive for markedly reduced activity but may miss intermediate activity, particularly in heterozygous females.
• Quantitative spectrophotometric assay — reference method; reports enzyme activity (U/g Hb or U/10^12 RBC). Use s*x- and population-specific reference ranges.
• Cytochemical/flow cytometry assays — demonstrate mosaicism and percentage of deficient cells; useful for identifying heterozygous females.
• Molecular/genetic testing — definitive for genotype; recommended when enzyme tests are ambiguous, after transfusion, or for female carriers.
Interpretation caveats
• Low enzyme activity in a male or clearly low value → consistent with G6PD deficiency (correlate clinically).
• Normal enzyme during/shortly after hemolysis or post-transfusion → does not exclude deficiency; repeat testing after recovery or perform genetic testing.
• Females with normal/borderline enzyme → consider heterozygosity; use cytochemical testing or molecular testing for confirmation.
• Always report method, units, reference range, and any preanalytic issues (recent hemolysis, transfusion, newborn status).
Tips
• If hemolysis is obvious and urgent: treat clinically now; send G6PD assay but plan repeat testing after recovery or order molecular testing.
• Do not rely on a single normal enzyme test in a recently transfused or actively hemolyzing patient.
• For females with suspicious history or borderline enzyme results: pursue cytochemical or molecular testing.
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