This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by de Moraes, R. R.
Right arrow Articles by Funderburk, J. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by de Moraes, R. R.
Right arrow Articles by Funderburk, J. E.
Agricola
Right arrow Articles by de Moraes, R. R.
Right arrow Articles by Funderburk, J. E.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, June 1999, p. 2307-2311, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Methods for Detection of Anticarsia gemmatalis Nucleopolyhedrovirus DNA in Soildagger

R. R. de Moraes,1,Dagger J. E. Maruniak,1,* and J. E. Funderburk2

Entomology and Nematology Department, University of Florida, Gainesville, Florida 32611,1 and Entomology and Nematology Department and North Florida Research and Education Center, University of Florida, Quincy, Florida 323512

Received 21 December 1998/Accepted 19 March 1999

Two methods, phenol-ether and magnetic capture-hybridization (MCH), were developed and compared with regard to their sensitivities and abilities to extract the DNA of the insect baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) from soil and to produce DNA amplifiable by PCR. Laboratory experiments were performed with 0.25 g of autoclaved soil inoculated with different viral concentrations to optimize both methods of baculovirus DNA extraction and to determine their sensitivities. Both procedures produced amplifiable DNA; however, the MCH method was 100-fold more sensitive than the phenol-ether procedure. The removal of PCR inhibitors from the soil appeared to be complete when MCH was used as the viral DNA isolation method, because undiluted aliquots of the DNA preparations could be amplified by PCR. The phenol-ether procedure probably did not completely remove PCR inhibitors from the soil, since PCR products were observed only when the AgMNPV DNA preparations were diluted 10- or 100-fold. AgMNPV DNA was detected in field-collected soil samples from 15 to 180 days after virus application when the MCH procedure to isolate DNA was coupled with PCR amplification of the polyhedrin region.


* Corresponding author. Mailing address: Entomology and Nematology Department, University of Florida, P.O. Box 110620, Gainesville, FL 32611. Phone: (352) 392-1901. Fax: (352) 392-0190. E-mail: marun{at}nervm.nerdc.ufl.edu.

dagger Publication no. R-06242 of the Florida Agricultural Experiment Station.

Dagger Present address: DuPont Ag Enterprises, Stine-Haskell Research Center, Newark, DE 19714.


Applied and Environmental Microbiology, June 1999, p. 2307-2311, Vol. 65, No. 6
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Yankson, K. K., Steck, T. R. (2009). Strategy for Extracting DNA from Clay Soil and Detecting a Specific Target Sequence via Selective Enrichment and Real-Time (Quantitative) PCR Amplification. Appl. Environ. Microbiol. 75: 6017-6021 [Abstract] [Full Text]  
  • Thorne, C. M., Otvos, I. S., Conder, N., Levin, D. B. (2007). Development and Evaluation of Methods To Detect Nucleopolyhedroviruses in Larvae of the Douglas-Fir Tussock Moth, Orgyia pseudotsugata (McDunnough). Appl. Environ. Microbiol. 73: 1101-1106 [Abstract] [Full Text]