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Volume 32, Number 4, July-August 2001
Page(s) 363 - 370
DOI: 10.1051/apido:2001136

Apidologie 32 (2001) 363-370

Development of a fast and reliable diagnostic method for American foulbrood disease (Paenibacillus larvae subsp. larvae) using a 16S rRNA gene based PCR

Wim Dobbelaerea, b, Dirk C. de Graafa and Johan E. Peetersa, b

a  Veterinary and Agrochemical Research centre, Groeselenberg 99, 1180 Brussels, Belgium
b  University of Ghent, Laboratory of Zoophysiology, Krijgslaan 281 S33, 9000 Ghent, Belgium

(Received 18 January 2001; revised 9 April 2001; accepted 1 May 2001)

Paenibacillus larvae larvae is the causative agent of American foulbrood (AFB), a serious honeybee disease. The ability to detect and identify this organism promptly is important to allow effective sanitary measures in case of AFB. P. larvae larvae is a fastidious and slow-growing bacterium and primary isolation and presumptive identification can be difficult and time-consuming. In this study the use of PCR is described for a rapid and reliable diagnosis. The developed PCR assay is specific for P. larvae as no amplicons were produced from 13 related or hive-related species. Only with P. larvae larvae and P. larvae pulvifaciens DNA as template, an amplicon was formed. A positive reaction was also observed when DNA was extracted directly from remains of an AFB diseased larva. Thus this PCR assay provides a reliable diagnosis for AFB in only 4 hours.

Key words: American foulbrood / diagnosis / Paenibacillus larvae larvae / pcr / 16s rRNA

Correspondence and reprints: Wim Dobbelaere

© INRA, EDP Sciences, DIB, AGIB 2001