Improved method for the detection of Paenibacillus larvae subsp. larvae, the cause of American Foulbrood of honey bees, by using polymerase chain reaction with subspecies specific primers

A. M. ALIPPI, A. C. LÓPEZ, AND O. MARIO AGUILAR

LETTERS IN APPLIED MICROBIOLOGY

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2004 vol. 39 p. 25 - 33

0266-8254

Aims: A reliable procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of AmericanFoulbrood disease of honey bees (Apis mellifera L.) based on the polymerase chain reaction (PCR) and subspecies –specific primers is described.Methods and Results: By using ERIC-PCR, an amplicon of ca 970 bp was found among P. l. larvae strainsbut not in other closely related species. Based on the nucleotide sequence data of this amplicon, we designed the pairof oligonucleotides KAT 1 and KAT 2, which were assayed as primers in a PCR reaction. A PCR amplicon ofthe expected size ca 550 bp was only found in P. l. larvae strains.Conclusions: This PCR assay provides a specific detection for P. l. larvae.Significance and Impact of the Study: The developed PCR assay is highly specific because can differentiatePaenibacillus larvae subsp. larvae from the closely related Paenibacillus larvae subsp. pulvifaciens. The techniquecan be directly used to detect presence or absence of P. l. larvae spores in honey bee brood samples andcontaminated honeys.