Investigations

1st investigations to order

peripheral blood smear

Test
Result
Test

Macrocytic anaemia and hypersegmented neutrophils are classically seen in folate and vitamin B12 (cobalamin) deficiencies. Macrocytes are seen in early folate deficiency, but they can also be present in other conditions.[Figure caption and citation for the preceding image starts]: Megaloblastic macrocytic anaemia: A. Peripheral blood smear of a patient with megaloblastic anaemia. B. Peripheral blood smear of healthy individualPhotomicrograph from Mark J. Koury, MD; used with permission [Citation ends].com.bmj.content.model.Caption@3958de5

Result

macrocytosis, anisocytosis, poikilocytosis, hypersegmented neutrophils

FBC

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Result
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Elevated mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) can precede anaemia by a few weeks; thrombocytopenia and neutropenia are seen in advanced cases.[40] Can be seen in other conditions. Not useful to rule out folate deficiency.

Result

low haemoglobin, elevated MCV and MCH; increased MCV and MCH may be absent or less than expected in combined folate and iron deficiency; thrombocytopenia, neutropenia

reticulocyte count

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Result
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Low count indicates decreased production. Can be present in other deficiency states or marrow failure disorders.

Result

low corrected reticulocyte count

Investigations to consider

serum folate

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Result
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Earliest indicator of folate deficiency. Performed as initial screening test. May not detect the 5% of patients who have folate deficiency but normal serum folate levels.[55][56]

Result

low

red blood cell folate

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Result
Test

Better indicator of tissue folate status than serum folate.[56] Can help detect the 5% of folate deficient patients with normal serum folate.

May be normal in acute folate deficiency.

Red blood cell folate is low in >50% of patients with vitamin B12 (cobalamin) deficiency.

A more complex and expensive test than serum folate.

Result

low

serum vitamin B12

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Result
Test

Extremely important test.

Underlying vitamin B12 (cobalamin) deficiency should be ruled out before implementing therapy with folic acid, because such therapy may mask neurological complications of untreated vitamin B12 deficiency.

Vitamin B12 deficiency and folate deficiency can co-exist in certain patients.

Result

normal

serum LDH

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Result
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Sign of ineffective erythropoiesis, present in advanced anaemia.

Not useful to rule in or rule out folate deficiency.

Result

elevated

serum unconjugated bilirubin

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Result
Test

Sign of ineffective erythropoiesis, present in advanced anaemia.

Not useful to rule in or rule out folate deficiency.

Result

elevated

serum iron panel

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Result
Test

Sign of ineffective erythropoiesis.

Not useful to rule in or rule out folate deficiency.

Result

elevated serum iron, ferritin, transferrin

plasma or serum methylmalonic acid

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Result
Test

Levels rise in vitamin B12 (cobalamin) deficiency but are normal in folate deficiency.[58][59]

Level affected by renal function.

Result

normal

plasma homocysteine

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Result
Test

Useful in borderline folate levels, suspected combined deficiency states, and folate-deficient patients with normal serum folate.[55]

Elevated in >95% of patients with vitamin B12 (cobalamin) deficiency.

Sensitivity is only 86% in detecting clinical folate deficiency states.[58]

Expensive.

Level affected by renal function.

Result

elevated

bone marrow aspirate/biopsy

Test
Result
Test

Classic findings in both folate and vitamin B12 (cobalamin) deficiencies.

Not required to confirm diagnosis.[Figure caption and citation for the preceding image starts]: Megaloblastic marrow cellsPhotomicrograph from Mark J. Koury, MD; used with permission [Citation ends].com.bmj.content.model.Caption@7cbfd35b

Result

megaloblastic changes, erythroid hyperplasia, abnormal nuclear appearance

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