Hematology · PANCE / PANRE

G6PD Deficiency

X-linked enzyme deficiency causing episodic oxidative hemolysis in response to drugs, infection, or fava beans.

Also known as: G6PD, glucose-6-phosphate dehydrogenase deficiency, favism

Overview

Inherited deficiency of glucose-6-phosphate dehydrogenase, the rate-limiting enzyme of the hexose monophosphate (pentose phosphate) shunt. Reduces NADPH availability, impairing red cell defenses against oxidative stress and predisposing to acute hemolysis under oxidant exposure.

Epidemiology

Most common enzymopathy worldwide, affecting ~400 million people. X-linked recessive — males more severely affected. Highest prevalence in African (G6PD A-, ~10% of US Black males), Mediterranean, Middle Eastern, and Southeast Asian populations. Geographic overlap with malaria endemicity (heterozygote advantage).

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Question 1HematologyEasy
A 28-year-old African American man is started on primaquine for malaria prophylaxis before international travel. Two days later he develops fatigue, scleral icterus, and dark urine. Laboratory studies show a hemoglobin of 8.4 g/dL, an elevated reticulocyte count, indirect hyperbilirubinemia, and bite cells with occasional Heinz bodies on the peripheral smear. Which of the following is the most likely diagnosis?
  • ASickle cell disease
  • BAutoimmune hemolytic anemia
  • CCold agglutinin disease
  • DG6PD deficiency
Reveal answer & full explanation
Correct answer: D — G6PD deficiency
  • ASickle cell disease
  • BAutoimmune hemolytic anemia
  • CCold agglutinin disease
  • DG6PD deficiency

Why G6PD deficiency is correct

  • Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked recessive defect of the hexose monophosphate shunt that leaves red cells unable to regenerate glutathione and defend against oxidative stress.
  • An oxidant exposure - here primaquine (also dapsone, sulfonamides, nitrofurantoin, fava beans, or infection) - triggers acute hemolysis within hours to days, producing anemia, indirect hyperbilirubinemia, and hemoglobinuria (dark urine).
  • The peripheral smear shows bite cells and Heinz bodies (precipitated, denatured hemoglobin), and the African American background fits the common A- variant.
  • Management is to stop the offending agent with supportive care; screen for G6PD before prescribing oxidant antimalarials.

Why the others are wrong

  • Sickle cell disease — Causes vaso-occlusive pain from lifelong HbS polymerization, not an acute hemolytic episode newly triggered by a drug, and the smear shows sickled cells rather than bite cells and Heinz bodies (anchoring on the patient's race).
  • Autoimmune hemolytic anemia — Warm AIHA is Coombs-positive with spherocytes and is not precipitated by an oxidant drug; the oxidative smear findings point elsewhere (premature closure on 'hemolytic anemia').
  • Cold agglutinin disease — Cold agglutinin hemolysis follows cold exposure or Mycoplasma/EBV infection with red-cell agglutination on the smear, not an oxidant drug exposure (buzzword-matching hemolysis without the trigger).
Question 2HematologyEasy
A 25-year-old man of Mediterranean ancestry develops severe hemolytic anemia (Hgb 6.2 g/dL) and hemoglobinuria 2 days after starting dapsone for Pneumocystis prophylaxis. Peripheral blood smear (PBS) shows bite cells and Heinz bodies on crystal violet stain. LDH is markedly elevated. Which of the following is the most likely diagnosis?
  • AHereditary spherocytosis
  • BParoxysmal nocturnal hemoglobinuria
  • CAutoimmune hemolytic anemia
  • DG6PD deficiency
Reveal answer & full explanation
Correct answer: D — G6PD deficiency
  • AHereditary spherocytosis
  • BParoxysmal nocturnal hemoglobinuria
  • CAutoimmune hemolytic anemia
  • DG6PD deficiency

Why G6PD deficiency is correct

  • Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme deficiency worldwide.
  • X-linked recessive — affects males predominantly; female carriers are rarely symptomatic.
  • Pathophysiology: G6PD generates NADPH, which maintains reduced glutathione that protects RBCs from oxidative stress; when deficient, oxidant stress denatures hemoglobin, which precipitates as Heinz bodies, driving both extravascular (splenic) and intravascular hemolysis.
  • Triggers: oxidant drugs (primaquine, dapsone, nitrofurantoin, rasburicase, sulfamethoxazole), infections, and fava beans.
  • Peripheral blood smear (PBS) findings: bite (blister) cells — macrophages pluck out Heinz bodies — and Heinz bodies on crystal violet stain.
  • Treatment: remove the offending agent, supportive care, and transfusion for severe anemia.

Why the others are wrong

  • Autoimmune hemolytic anemia — has a positive direct Coombs and is not oxidant-drug triggered (confused-with an antibody-mediated hemolysis).
  • Paroxysmal nocturnal hemoglobinuria — also causes intravascular hemolysis with hemoglobinuria, but it is complement-mediated from loss of GPI-anchored CD55/CD59 (diagnosed by flow cytometry), is not triggered by dapsone, and does not produce Heinz bodies or bite cells (anchoring on hemoglobinuria).
  • Hereditary spherocytosis — a chronic inherited membrane defect with spherocytes, elevated MCHC, and a positive EMA-binding or osmotic fragility test, not an acute crisis provoked by an oxidant drug; bite cells and Heinz bodies are absent (confused-with an inherited hemolytic anemia).

Additional high-yield points

  • G6PD assay timing: a quantitative assay can read falsely NORMAL during an acute episode because young reticulocytes carry higher enzyme activity; retest after recovery once the reticulocytosis has resolved rather than relying on a single fixed interval.
  • African A- variant is milder, with partial enzyme activity retained in younger RBCs; the Mediterranean variant tends to be more severe.
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Risk factors

  • Family history (X-linked)
  • African, Mediterranean, Middle Eastern, Southeast Asian ancestry
  • Triggers — oxidant exposure: dapsone, primaquine, tafenoquine, sulfonamides (TMP-SMX), nitrofurantoin, methylene blue, rasburicase, high-dose aspirin, naphthalene (mothballs), henna
  • Infection (most common trigger overall)
  • Diabetic ketoacidosis
  • Fava bean ingestion (Mediterranean variant)

Pathophysiology

G6PD produces NADPH, which reduces glutathione (GSH); reduced glutathione neutralizes peroxides and other oxidants. Without adequate NADPH/GSH, oxidant exposure causes denaturation of hemoglobin (Heinz bodies) and membrane damage, leading to intravascular and extravascular hemolysis. Mature RBCs cannot synthesize new enzyme, so older cells lyse first; reticulocytes have more enzyme and survive — hemolysis is self-limited in mild variants.

Clinical presentation

Symptoms

  • Acute hemolytic episode 24-72 hours after oxidant exposure: dark/cola-colored urine (hemoglobinuria), back/abdominal pain, jaundice, fatigue, dyspnea
  • Neonatal jaundice — especially Mediterranean variant; can be severe and cause kernicterus
  • Most patients asymptomatic between triggers
  • Severe variants (Mediterranean): chronic non-spherocytic hemolytic anemia

Signs / physical exam

  • Pallor and jaundice during hemolytic episode
  • Mild splenomegaly possible
  • Dark urine (hemoglobinuria)
  • Neonatal: scleral icterus, jaundice in first week of life

Classic findings

Dark urine and jaundice 1-3 days after starting TMP-SMX (or dapsone, or fava bean ingestion) in a young man of African or Mediterranean descent.

Differential diagnosis

  • Autoimmune hemolytic anemia — Positive direct Coombs (DAT), spherocytes; no oxidative trigger
  • Hereditary spherocytosis — Chronic mild hemolysis, spherocytes, positive osmotic fragility/EMA binding; family history
  • Pyruvate kinase deficiency — Chronic non-spherocytic hemolytic anemia; PK enzyme assay
  • Sickle cell disease — Sickle cells on smear, hemoglobin electrophoresis; G6PD can coexist and worsen crises
  • Drug-induced immune hemolysis — Positive DAT, temporally related to drug, mechanism distinct from oxidant injury
  • TTP/HUS — Schistocytes, thrombocytopenia, end-organ injury

Diagnostic workup

Diagnostic criteria

Low quantitative G6PD enzyme activity in a stable patient (away from acute hemolytic episode). Newborn screening available in many states.

Labs

  • CBC — normocytic anemia, reticulocytosis (peaks ~7 days after trigger)
  • Peripheral smear — bite cells (membrane removed by splenic macrophages around denatured Hb), blister cells, Heinz bodies (require supravital stain such as crystal violet)
  • Markers of hemolysis: elevated LDH, indirect hyperbilirubinemia, low haptoglobin, hemoglobinuria on UA
  • G6PD enzyme activity assay — diagnostic; quantitative or qualitative (fluorescent spot test)
  • TIMING CAVEAT: G6PD level may be falsely normal during acute hemolysis (older deficient cells already lysed; reticulocytes have higher enzyme activity). Repeat 2-3 months after recovery
  • Direct Coombs negative (rules out autoimmune hemolysis)

Imaging

  • Not routinely indicated

Diagnostic algorithm

flowchart TD
  A[Oxidant exposure<br/>TMP-SMX, dapsone, primaquine,<br/>fava beans, infection] --> B[Oxidant stress on RBC]
  B --> C{Adequate NADPH<br/>from G6PD?}
  C -->|Yes - normal| D[Glutathione reduces<br/>oxidants — no hemolysis]
  C -->|No - G6PD deficient| E[Hemoglobin denaturation<br/>→ Heinz bodies]
  E --> F[Spleen removes<br/>Heinz bodies<br/>→ bite cells]
  E --> G[Membrane damage<br/>→ intravascular<br/>hemolysis]
  G --> H[Hemoglobinuria<br/>↑LDH, ↓haptoglobin,<br/>indirect ↑bili]
  F --> I[Acute anemia<br/>24-72h after trigger]
  G --> I
  I --> J[Reticulocytosis,<br/>self-limited in<br/>African A- variant]
G6PD deficiency hemolytic cascade — oxidant trigger to clinical hemolysis.

Treatment

First-line

  • Acute hemolysis: identify and remove offending agent immediately
  • Supportive care — IV fluids to maintain renal perfusion and clear hemoglobinuria
  • Transfusion for severe symptomatic anemia or hemodynamic instability
  • Prevention is primary: educate patient about drugs and foods to avoid; provide list of contraindicated medications
  • Neonatal jaundice: phototherapy, exchange transfusion if severe (to prevent kernicterus)

Second-line / adjunct

  • Screen for G6PD deficiency BEFORE prescribing dapsone, primaquine, tafenoquine, rasburicase in at-risk populations
  • Avoid in family members; X-linked inheritance counseling
  • Folic acid for chronic hemolytic variants
  • Splenectomy rarely indicated (chronic non-spherocytic severe variants only)

Complications

  • Acute renal failure from hemoglobinuria (pigment nephropathy) in severe hemolysis
  • Kernicterus in neonates with severe hyperbilirubinemia
  • Cholelithiasis (pigment stones) from chronic hemolysis
  • Drug-induced fatality if oxidant given despite known deficiency (especially rasburicase, dapsone)
  • Anemia-related complications during pregnancy and intercurrent illness

PANCE pearls

  • Bite cells and Heinz bodies are the hematologic signature of oxidative hemolysis. Heinz bodies require supravital staining.
  • G6PD enzyme level can be FALSELY NORMAL during acute hemolysis — older deficient cells are gone, reticulocytes are enzyme-rich. Always confirm with a repeat assay 2-3 months later.
  • Rasburicase is absolutely contraindicated in G6PD deficiency — causes severe acute hemolysis and methemoglobinemia. Screen before use, especially in tumor lysis prophylaxis.
  • Female heterozygotes may have variable phenotype due to X-inactivation (lyonization); some experience clinically significant hemolysis.
  • Hemolytic anemia 24-72 hours after starting TMP-SMX, dapsone, primaquine, or eating fava beans is the classic vignette.
  • Fava bean reactions (favism) occur primarily in Mediterranean variant, not African A- variant.
  • African (A-) variant: hemolysis is self-limited because reticulocytes have near-normal enzyme. Mediterranean variant: enzyme deficient even in reticulocytes, hemolysis can be severe and protracted.

References

  • WHO 2022 — WHO Technical specifications series for G6PD testing
  • Cappellini & Fiorelli — Glucose-6-phosphate dehydrogenase deficiency (Cappellini & Fiorelli, Lancet 2008)
  • Luzzatto et al. — Favism and Glucose-6-Phosphate Dehydrogenase Deficiency (NEJM 2018)
  • Beutler — G6PD deficiency (Beutler, Blood 1994)

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