Intrinsic Factor Blocking Antibody, Serum
Confirming the diagnosis of pernicious anemia
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
The cobalamins, also referred to as vitamin B12, are a group of closely related enzymatic cofactors involved in the conversion of methylmalonyl-coenzyme A to succinyl-coenzyme A and in the synthesis of methionine from homocysteine. Vitamin B12 deficiency can lead to megaloblastic anemia and neurological deficits. The latter may exist without anemia, or precede it. Adequate replacement therapy will generally improve or cure cobalamin deficiency. Unfortunately, many other conditions, which require different interventions, can mimic the symptoms and signs of vitamin B12 deficiency. Moreover, even when cobalamin deficiency has been established, clinical improvement may require different dosages or routes of vitamin B12 replacement, depending on the underlying cause. In particular, patients with pernicious anemia (PA), possibly the commonest type of cobalamin deficiency in developed countries, require either massive doses of oral vitamin B12 or parenteral replacement therapy. The reason is that in PA patients suffer from gastric mucosal atrophy, most likely caused by a destructive autoimmune process. This results in diminished or absent gastric acid, pepsin and intrinsic factor (IF) production. Gastric acid and pepsin are required for liberation of cobalamin from binding proteins, while IF binds the free vitamin B12, carries it to receptors on the ileal mucosa, and facilitates its absorption. Most PA patients have autoantibodies against gastric parietal cells or intrinsic factor, with the latter being very specific but only present in approximately 50% of cases. By contrast, parietal cell antibodies are found in approximately 90% of PA patients, but are also found in a significant proportion of patients with other autoimmune diseases, and in approximately 2.5% (4th decade of life) to approximately 10% (8th decade of life) of healthy individuals.
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
The aim of the work-up of patients with suspected vitamin B12 deficiency is to first confirm the presence of deficiency and then to establish its most likely etiology.
Measurement of serum vitamin B12, either preceded or followed by serum methylmalonic acid measurement, is the first step in diagnosing pernicious anemia (PA). If these tests support deficiency, then intrinsic factor blocking antibody (IFBA) testing is indicated to confirm PA as the etiology. A positive IFBA test supports very strongly a diagnosis of PA. Since the diagnostic sensitivity of IFBA testing for PA is only around 50%, an indeterminate or negative IFBA test does not exclude the diagnosis of PA. In these patients, either PA or another etiology, such as malnutrition, may be present. Measurement of serum gastrin levels will help in these cases. In patients with PA, fasting serum gastrin is elevated to >200 pg/mL in an attempted compensatory response to the achlorhydria seen in this condition.
For a detailed overview of the optimal testing strategies in PA diagnosis, see ACASM/83632 Pernicious Anemia Cascade, Serum, and associated Pernicious Anemia Testing Cascade in Special Instructions.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
Do not order intrinsic factor blocking antibody (IFBA) testing in patients who have received a vitamin B12 injection within the last 2 weeks. High free serum vitamin B12 levels, as may be seen within the first 2 weeks after a vitamin B12 injection, can interfere in the IFBA assay, leading to false-positive results. We reflex all positive IFBA tests that have not been ordered through the Pernicious Anemia Cascade to vitamin B12 measurement. If this yields a level >800 ng/L, we append a comment to the report indicating a possible false-positive result.
Some patients with other autoimmune diseases may have positive IFBA assays without suffering from pernicious anemia (PA). This is reported inparticular in patients with autoimmune thyroid disease or type I diabetes mellitus. In the validation of this assay, 24 individuals with these autoimmune endocrine diseases were tested, and all were IFBA negative. However, 5 of 15 of patients with rheumatoid arthritis were IFBA positive during the validation of this assay. The literature suggests such individuals may in fact be at risk of later development of PA.
Since this is a competitive assay, the risk of heterophile antibody interference is low. During validation, 24 human anti-mouse antibody positive specimens and 25 specimens with other heterophile antibodies were tested and all were IFBA negative. However, if the clinical picture does not agree with the IFBA test result, the laboratory should be consulted for advice.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Toh BH, Van Driel IR, Gleeson PA: Pernicious anemia. N Engl J Med 1997;337:1441-1448
2. Klee GG: Cobalamin and folate evaluation: measurement of methylmalonic acid and homocysteine vs vitamin B12 and folate. Clin Chem 2000;46:1277-1283
3. Ward PC: Modern approaches to the investigation of vitamin B12 deficiency. Clin Lab Med 2002:22;435-445
4. Stabler SP, Allen RH: Vitamin B12 deficiency as a worldwide problem. Ann Rev Nutr 2004;24:299-326