DETECTION AND IDENTIFICATION OF MULTIPLE BACULOVIRUSES USING THE POLYMERASE CHAIN-REACTION (PCR) AND RESTRICTION-ENDONUCLEASE ANALYSIS

A technique using the polymerase chain reaction (PCR) and restriction analysis was developed for the simultaneous detection of eight baculov iruses. The baculoviruses detected by this technique were Autographa c alifornica multiple-embedded nuclear polyhedrosis virus (MNPV), Antica rsia gemmatalis MNPV, Bombyx mori MNPV, Orgyia pseudotsugata MNPV. Spo doptera frugiperda MNPV, S. exigua MNPV, Anagrapha falcifera MNPV, Hel iothis zea single-embedded nuclear polyhedrosis virus (SNPV). A highly conserved DNA sequence within the coding region of the polyhedrin gen e was targeted for amplification. One pair of degenerate primers was d esigned, and PCR conditions were optimized to produce 575 base pair fr agments For all eight baculoviruses. Restriction analysis of the PCR p roducts resulted in distinct profiles for each virus. This technique w ould be useful in monitoring the release of wild type as well as genet ically engineered baculoviruses. Copyright (C) 1997 Elsevier Science B .V.