Mnemonic

B12 and Folate Testing

A memory aid for investigating suspected B12 and folate deficiency.

Expansion

Serum levels, then the metabolites, then the cause

Expansion

Initial tests: serum B12 and serum folate, alongside the full blood count and blood film.

Film findings: macrocytosis, oval macrocytes, hypersegmented neutrophils (more than 5 per cent with five lobes, or any with six), and in severe cases pancytopenia with ineffective erythropoiesis, giving a raised LDH and bilirubin that can mimic haemolysis.

Pitfalls of the serum B12 assay

  • Falsely low: pregnancy, oral contraceptives, folate deficiency itself, myeloma, metformin, and in about half of people with a low result there is no true deficiency
  • Falsely normal despite deficiency: liver disease, myeloproliferative disease, and small intestinal bacterial overgrowth
  • The active (holotranscobalamin) assay is more specific

Second line metabolites, where the result is borderline or discordant with the clinical picture

B12 deficiency Folate deficiency
Methylmalonic acid Raised Normal
Homocysteine Raised Raised

So a raised methylmalonic acid is the marker that separates them.

Finding the cause of B12 deficiency: intrinsic factor antibodies are specific but only about 50 per cent sensitive; parietal cell antibodies are sensitive but non-specific. Consider pernicious anaemia, gastrectomy, ileal disease or resection, Crohn’s, metformin, proton pump inhibitors, and a vegan diet.

Neurological disease can occur with a normal haemoglobin and a normal MCV, which is why B12 must be checked in unexplained neuropathy, cognitive decline or ataxia, and why treatment should not wait for anaemia.