Development of a highly sensitive direct microtiter plate ELISA for human serum testosterone |
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Authors: | M Banerjee S Bhattacharyya C Das |
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Institution: | (1) Department of Biochemistry, All India Institute of Medical Sciences, 110 029 New Delhi;(2) Department of Biochemistry, Santosh World Medical Academy, Ghaziabad, U.P. |
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Abstract: | A simple, highly sensitive, direct, competitive ELISA for human serum testosterone has been indigenously developed. Specific
antisera against testosterone were raised in rabbits using testosterone—3carboxymethyl oxime (CMO)—bovine serum albumin (BSA)
as the antigen. For the enzyme conjugate, testoterone—3CMO was coupled with horse raddish peroxidase by the active ester method.
The standard curve covered a wide range from 3.9 pg/ml to 500 pg/ml. The inter and intra-assay variation were found to be
low and within the acceptable limits. Specificity and accuracy for the assay was established by having negligible crossreactivity
with the related steroids and an excellent parallelism between the sample and standard dilution curve. Samples measured by
RIA and ELISA showed very high degree of correlation (r=0.991). |
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Keywords: | Testosterone Direct ELISA Serum testosterone Sensitive ELISA |
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