A PCR-based method that permits specific detection of Paenibacillus larvae subsp. larvae, the cause of American Foulbrood of honey bees, at the subspecies level.
OBJECTIVE
A reliable procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of American Foulbrood disease of honey bees (Apis mellifera L.) based on the polymerase chain reaction (PCR) and subspecies - specific primers is described.


RESULTS
By using ERIC-PCR, an amplicon of ca 970 bp was found among P. l. larvae strains but not in other closely related species. Based on the nucleotide sequence data of this amplicon, we designed the pair of oligonucleotides KAT 1 and KAT 2, which were assayed as primers in a PCR reaction. A PCR amplicon of the 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.


CONCLUSIONS
The developed PCR assay is highly specific because can differentiate Paenibacillus larvae subsp. larvae from the closely related Paenibacillus larvae subsp. pulvifaciens. The technique can be directly used to detect presence or absence of P. l. larvae spores in honey bee brood samples and contaminated honeys.
Alippi
A M
AM
Facultad de Ciencias Agrarias y Forestales, Centro de Investigaciones de Fitopatología (CIDEFI), Universidad Nacional de La Plata, La Plata, Argentina. alippi@biol.unlp.edu.ar
López
A C
AC
Aguilar
O M
OM
D000818:Animals; D001406:Bacillaceae / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D001516:Bees / Q000254:growth & development / Q000382:microbiology*; D017931:DNA Primers; D006722:Honey / Q000382:microbiology; D007814:Larva / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D013171:Spores, Bacterial / Q000235:genetics
D017931:DNA Primers