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Applied and Environmental Microbiology, June 1999, p. 2307-2311, Vol. 65, No. 6
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.
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Methods for Detection of Anticarsia
gemmatalis Nucleopolyhedrovirus DNA in Soil

*
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.
Publication no. R-06242 of the Florida Agricultural Experiment Station.
Present address: DuPont Ag Enterprises, Stine-Haskell Research
Center, Newark, DE 19714.
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