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 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.
de Moraes
R R
RR
Entomology and Nematology Department, University of Florida, Gainesville 32611, USA.
Maruniak
J E
JE
D000818:Animals; D012831:Bombyx / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D009036:Moths / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping; D018411:Spodoptera / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral; C027908:endodeoxyribonuclease DdeI; D015252:Deoxyribonucleases, Type II Site-Specific; C037130:GCGC-specific type II deoxyribonucleases; C024400:GTYRAC-specific type II deoxyribonucleases