Issue |
Apidologie
Volume 32, Number 4, July-August 2001
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Page(s) | 363 - 370 | |
DOI | https://doi.org/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, ba 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)
Abstract
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
e-mail: widob@var.fgov.be
© INRA, EDP Sciences, DIB, AGIB 2001