BIOLUMINOMETRIC METHOD FOR REAL-TIME DETECTION OF REVERSE-TRANSCRIPTASE ACTIVITY

A simple and sensitive technique for detection of reverse transcriptas e (RT) activity in real time has been developed. The technique is base d on continuous detection of the inorganic pyrophosphate formed in the RT-catalyzed reaction by a luminometric method. The technique has bee n used for continuous monitoring of RT-catalyzed DNA synthesis on both homo- and heteropolymeric templates. The assay is sensitive and yield s linear responses between 1.5-960 mU of avian myeloblastosis virus RT (AMB-RT). The assay was used for detection of the inhibitory effect o f dideoxythymidine (ddTTP) on tile AMV-RT activity and also for real-t ime detection of single-base incorporation events catalyzed by AMV-RT. The possibility of using the new technique for other applications is discussed.