Adaptation of the Troponin T ELISA Test to a Microplate Immunoassay Reader

Abstract

Summary: Troponin T concentration in serum is usually measured by the automated method developed by Boehringer Mannheim for the ES-series of analysers. These instruments need at least 140 μΐ of serum and 700 μΐ of reagents for a single analysis, which takes 90 min. We describe an alternative procedure, using streptavidin-coated microtitre plates, troponin T reagents of Boehringer Mannheim and an ELIS Α-reader to measure the concentration of troponin T. The present assay needs only 30 μΐ of sample and 200 μΐ of reagents, and it takes 75 min; the detection limit is 0.10 μg/l. We also assessed the microtitre plate method for sensitivity and precision and compared the results with those measured by an ES-300 automatic analyser. Both methods have the same measurement range for troponin T of 0.1 to 15 μg/l. For daily routine use of the microtitre plate method we recommend duplicate determinations.

Comparison of the Troponin T and Troponin I ELISA Tests, as Measured by Microplate Immunoassay Techniques, in Diagnosing Acute Myocardial Infarction

“…3). The between series variation of troponin T was smaller than in the first modification (14), after addition of a microcomputer with the Genesis® program for the calculation of the results. The between series variation of troponin I was below 10.4% (tab.…”

“…No. 1556428) produced by Boehringer Mannheim were used (14). Instead of the original tubes, the reaction was performed in the 250 μΐ wells of a microtitre plate after coating the wells with streptavidine.…”

“…Coated microtitre plates were stored at + 4 °C until used. Otherwise, the method was performed in a similar way to that presented earlier (14) with minor modifications (15). Standard, control and serum (30 μΐ each) were dispensed into the wells of the plate with a manual Proline 5-50 pipette of Biohit Co. (Helsinki, Finland).…”

“…In this article we describe an improved modification of our ELISA procedure (14) for measuring troponin T using a standard microtitre plate immunoassay reader (15) and an ELISA procedure for troponin I. For comparison, creatine kinase MB 1 ) mass was measured in serum.…”

“…This formed part of a larger study of patients coming to the hospital with chest pain. For this purpose we tried to reduce the between-series variation of our troponin T procedure (14). Secondly we studied the use of troponin I in the diagnosis of acute myocardial infarction using a standard microtitre immunoassay technique.…”