[Study on evaluation of human immunodeficiency virus reverse transcriptase inhibitor by ELISA].
OBJECTIVE
to establish an ELISA method for detecting the activity of reverse transcriptase.


METHODS
Oligo (dT) 15 was immobilized via its 5'-terminal phosphate to Covalink-NH microtiter plates,The biotin-dUTP was incorporated by reverse transcriptase. The products were detected and quantified using a colorimetric streptavidin-alkaline phosphatase reporter system.


RESULTS
We established an ELISA RT assay method and tested the inhibition activity of PFA and NVP respectively. Using this method we also studied the kinetic mechanism of AMV-RT and HIV-1 RT. At the same time, we evaluated the stability, repeatability, specificity and sensitivity of this ELISA method.


CONCLUSIONS
This ELISA method has particular advantages, such as simplicity, rapidity, specificity and little pollution, and is suitable for screening and studying of specific inhibitors of HIV-1 reverse transcriptase and other RTs.
Guo
Ying
Y
Department of Chemical Biology, School of Pharmaceutical Science, State Key laboratory of Natural and Biomimetic Drugs, Peking University, Beijing 100083, China.
Wang
Xiao-wei
XW
Xu
Yang
Y
Lv
Xiao
X
Wang
Rui-ping
RP
Liu
Jun-yi
JY
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000201:enzymology*
C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase