Abstract：
:A colorimetric reverse transcriptase assay (cRT assay) was developed for quantitative detection of HIV-1. In this format, reverse transcriptase incorporates biotin-labeled dUTP onto oligo-dT primers hybridized to poly A templates. The templates are covalently bound to the surface of microtiter wells. The amount of incorporated biotin-labeled dUTP is measured by binding horseradish peroxidase conjugated streptavidin, washing away unbound peroxidase, adding colorimetric substrate and then reading with a standard colorimetric reader. The sensitivity of the assay is very good. As little as 3 x 10(5) molecules of recombinant HIV-RT can be detected after 20 h of reaction time. Direct comparison using 3 cultured clinical isolates indicates that this level of detection is equivalent to the commercially available p24 antigen capture assay and the HIV-RNA assay based on branched DNA signal amplification. Other retroviruses, such as HIV-2 and feline immunodeficiency virus (FIV), can also be detected in this format. This non-isotopic assay is easy to perform and could provide a convenient and quantitative method for HIV study by monitoring reverse transcriptase, an essential activity in the infection process.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Suzuki K,Saito T,Kondo M,Osanai M,Watanabe S,Kano T,Kano K,Imai Mdoi
10.1016/0166-0934(95)00073-5subject
Has Abstractpub_date
1995-11-01 00:00:00pages
347-56issue
3eissn
0166-0934issn
1879-0984pii
0166093495000735journal_volume
55pub_type
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