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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.4/1.5


TITLE: RETROVIRUS DETECTION BY ELISA


In the ELISA, digoxigenin- and biotin-labeled desoxyuridine-triphosphates are incorporated in the presence of viral reverse transcriptase (RT) during the synthesis of a DNA strand along a synthetic target of single-stranded DNA molecules. The newly synthesized DNA is trapped on streptavidin-coated microtiter plates and subsequently detected immunologically by binding of peroxidase-labeled anti-digoxigenin antibodies and visualized by color development and finally quantified by measuring the bsorbance with an ELISA reader (1). Two different buffers are used to detect Mg2+-dependent RT as well as Mn2+-dependent RT of C-type retroviruses.

References:

1. Eberle J, Seibl R: A new method for measuring reverse transcriptase activity by ELISA. J Virol Meth 40: 347-356 (1992).


Guidelines prepared for CABRI by CERDIC, DSMZ, ECACC, INRC, November 1998
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