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Applied and Environmental Microbiology, May 1999, p. 2243-2245, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A PCR Detection Method for Rapid Identification of Paenibacillus larvae

V. A. Govan,1 M. H. Allsopp,2 and S. Davison1,*

Department of Microbiology, University of the Western Cape, Bellville 7535,1 and Plant Protection Research Institute, Agricultural Research Council, Stellenbosch 7599,2 South Africa

Received 29 October 1998/Accepted 2 March 1999

American foulbrood is a disease of larval honeybees (Apis mellifera) caused by the bacterium Paenibacillus larvae. Over the years attempts have been made to develop a selective medium for the detection of P. larvae spores from honey samples. The most successful of these is a semiselective medium containing nalidixic acid and pipermedic acid. Although this medium allows the growth of P. larvae and prevents the growth of most other bacterial species, the false-positive colonies that grow on it prevent the rapid confirmation of the presence of P. larvae. Here we describe a PCR detection method which can be used on the colonies that grow on this semiselective medium and thereby allows the rapid confirmation of the presence of P. larvae. The PCR primers were designed on the basis of the 16S rRNA gene of P. larvae and selectively amplify a 973-bp amplicon. The PCR amplicon was confirmed as originating from P. larvae by sequencing in both directions. Detection was specific for P. larvae, and the primers did not hybridize with DNA from closely related bacterial species.

* Corresponding author. Mailing address: Department of Microbiology, University of the Western Cape, Private Bag X17, Bellville 7535, South Africa. Phone: 27-21-9592216. Fax: 27-21-9592216. E-mail: Sean{at}mbiol.uwc.ac.za.

Applied and Environmental Microbiology, May 1999, p. 2243-2245, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Bakonyi, T., Derakhshifar, I., Grabensteiner, E., Nowotny, N. (2003). Development and Evaluation of PCR Assays for the Detection of Paenibacillus larvae in Honey Samples: Comparison with Isolation and Biochemical Characterization. Appl. Environ. Microbiol. 69: 1504-1510 [Abstract] [Full Text]  
  • Alippi, A. M., Lopez, A. C., Aguilar, O. M. (2002). Differentiation of Paenibacillus larvae subsp. larvae, the Cause of American Foulbrood of Honeybees, by Using PCR and Restriction Fragment Analysis of Genes Encoding 16S rRNA. Appl. Environ. Microbiol. 68: 3655-3660 [Abstract] [Full Text]  


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