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Applied and Environmental Microbiology, February 2007, p. 1101-1106, Vol. 73, No. 4
0099-2240/07/$08.00+0     doi:10.1128/AEM.01958-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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

Christine M. Thorne,1 Imre S. Otvos,2 Nicholas Conder,2 and David B. Levin3*

Department of Biology, University of Victoria, P.O. Box 3020 STN CSC, Victoria, British Columbia, Canada,1 Pacific Forestry Centre, 506 West Burnside Rd., Victoria, British Columbia V8Z 1M5, Canada,2 Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba R3T 5V6, Canada3

Received 17 August 2006/ Accepted 11 December 2006

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 (r2 = 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.

* Corresponding author. Mailing address: Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba R3T 5V6, Canada. Phone: (204) 474-7429. Fax: (204) 474-7512. E-mail: levindb{at}cc.umanitoba.ca.

{triangledown} Published ahead of print on 22 December 2006.

Applied and Environmental Microbiology, February 2007, p. 1101-1106, Vol. 73, No. 4
0099-2240/07/$08.00+0     doi:10.1128/AEM.01958-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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