Development and Evaluation of Methods To Detect Nucleopolyhedroviruses in Larvae of the Douglas-Fir Tussock Moth, Orgyia pseudotsugata (McDunnough)

Abstract

Various molecular methods are used to detect pathogenic microorganisms and viruses within their hosts, but these methods are rarely validated by direct comparison. Southern hybridization, enzyme-linked immunosorbent assay (ELISA), and a novel DNA extraction/PCR assay were used to detect Orgyia pseudotsugata multiple nucleopolyhedrovirus (OpMNPV) in Douglas-fir tussock moth larvae. PCR was more sensitive than Southern hybridization and ELISA at detecting semipurified virus. ELISA, however, was the most accurate method for detecting virus within larvae, given that Southern hybridization and PCR produced false-negative results (31% and 2.5%, respectively). ELISA may be preferable in some applications because virus infections can be quantified (r 2 ‫؍‬ 0.995). These results may be applicable to both applied and academic research that seeks to accurately identify the incidence of viruses and microorganisms that regulate insect populations.

Stability of Spodoptera litura nucleopolyhedrovirus in sodium dodecyl sulphate

“…Sodium dodecyl sulphate (SDS) is an anionic surfactant capable of disrupting non-covalent bonding in proteins and denaturing them with most proteins that lose their functions completely or partially in its presence (Yamada et al, 1998). In insect pathological works, SDS has been used routinely for extracting and purifying baculovirus polyhedra from infected cells of diseased insects for bioassays (Thorne et al, 2007). The compound lyses cells and expose the DNA.…”

Reovirus-Like Sequences Isolated from Adult Asian Citrus Psyllid, (Hemiptera: Psyllidae:Diaphorina citri)

“…Like most animals, insects are susceptible to diseases caused by viruses, and many viruses have been applied as insecticides (Tweeten et al 1981;Moscardi 1999). In the United States, Nuclear polyhedrosis viruses (NPVs) are used as insecticides (Thorne et al 2007). The NPVs suppress the corn earworm moth (Heliothis zea), the Asian gypsy moth (Lymantria dispar), and the Douglas fir tussock moth (Orgria pseudotsugata) (Shieh & Bohmfalk 1980).…”

An immunological method for granulovirus detection in larvae ofTuta absoluta: searching for isolates with prospects for biological control of this pest in Colombia

“…The detection range of samples obtained in this study (over 5 £ 10 4 OBs6 mL) is within the range reported by other authors for immunological techniques such as ELISA. For instance, Parola et al (2003) and Thorne et al (2007), using baculovirus as an antigen, were able to develop antibodies for the detection of granulovirus from insect larvae and complex samples of granulovirus formulations.…”