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Applied and Environmental Microbiology, May 2001, p. 2384-2387, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2384-2387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Acute Bee Paralysis Virus and Black Queen Cell Virus from Honeybees by Reverse Transcriptase PCR

Mongi Benjeddou, Neil Leat, Mike Allsopp,dagger and Sean Davison*

Department of Microbiology, University of the Western Cape, Bellville 7535, Cape Town, South Africa

Received 26 October 2000/Accepted 12 February 2001

A reverse transcriptase PCR (RT-PCR) assay was developed for the detection of acute bee paralysis virus (ABPV) and black queen cell virus (BQCV), two honeybee viruses. Complete genome sequences were used to design unique PCR primers within a 1-kb region from the 3' end of both genomes to amplify a fragment of 900 bp from ABPV and 700 bp from BQCV. The combined guanidinium thiocyanate and silica membrane method was used to extract total RNA from samples of healthy and laboratory-infected bee pupae. In a blind test, RT-PCR successfully identified the samples containing ABPV and BQCV. Sensitivities were approximately 1,600 genome equivalents of purified ABPV and 130 genome equivalents of BQCV.

* Corresponding author. Mailing address: Department of Microbiology, University of the Western Cape, Private Bag X17, Bellville 7535, Cape Town, South Africa. Phone: (2721) 959 2216. Fax: (2721) 959 2216. E-mail: sdavison{at}uwc.ac.za.

dagger Present address: Plant Protection Research Institute, Agricultural Research Council, Stellenbosch 7599, South Africa.

Applied and Environmental Microbiology, May 2001, p. 2384-2387, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2384-2387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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