Detection and identification of multiple baculoviruses using the polymerase chain reaction (PCR) and restriction...

A technique using the polymerase chain reaction (PCR) and restriction analysis was developed for the simultaneous detection of eight baculoviruses. The baculoviruses detected by this technique were Autographa californica multiple-embedded nuclear polyhedrosis virus (MNPV). Anticarsia gemmatalis MNPV, Bombyx mori MNPV, Orgyia pseudotsugata MNPV. Spodoptera frugiperda MNPV, S. exigua MNPV, Anagrapha falcifera MNPV, Heliothis zea single-embedded nuclear polyhedrosis virus (SNPV). A highly conserved DNA sequence within the coding region of the polyhedrin gene was targeted for amplification. One pair of degenerate primers was designed, and PCR conditions were optimized to produce 575 base pair fragments for all eight baculoviruses. Restriction analysis of the PCR products resulted in distinct profiles for each virus. This technique would be useful in monitoring the release of wild type as well as genetically engineered baculoviruses.