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

Paenibacillus larvae larvae is the causative agent of American foulbrood (A FB), a serious honeybee disease. The ability to detect and identify this or ganism promptly is important to allow effective sanitary measures in case o f AFB. P. larvae larvae is a fastidious and slow-growing bacterium and prim ary isolation and presumptive identification can be difficult and time-cons uming. In this study the use of PCR is described for a rapid and reliable d iagnosis. 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.