Increased sensitivity of detection of the blocking (type I) anti-intrinsic factor antibody.

Increasing the ratio of test serum to the intrinsic factor (IF) antigen should increase the sensitivity of anti-IF antibody assay, a useful, specific tool for diagnosing pernicious anemia. We therefore designed an assay in which this ratio was increased 25-fold over that of our current standard assay and 1,000-fold over that of most described methods. This assay was then compared with our standard assay, with one of the original assays, and with a commercial kit. Both of our assays, especially the newer sensitive method, detected antibody in much greater dilutions of positive serum than the other assays. The sensitive assay also detected antibody in 17% of pernicious anemia sera negative by our standard technique. We calculated that anti-IF antibody can be detected with the sensitive assay in 78% of our patients with pernicious anemia, compared with 73% by our standard assay, 65% by the commercial kit, and 61% by the original method of Gottlieb and colleagues. No antibody was detected in 35 control sera with the new assay, which suggests that the increased sensitivity did not come at the expense of diminished specificity. The increased sensitivity enhances the value of the anti-IF antibody as a major screening and diagnostic tool in pernicious anemia.