Detection of Paenibacillus larvae subspecies larvae spores in naturally infected bee larvae and artificially contaminated honey by PCR

摘要: American foulbrood (AFB), a severe bacterial disease of honeybee brood, has recently been found in Uruguayan apiaries. Detection the causative agent, Paenibacillus larvae subspecies larvae, is very important concern order to prevent dissemination and decrease honey production. Since spores are infective forms this pathogen, present work we report use polymerase chain reaction (PCR) detect P. l. subsp. from vitro cultures, with clinical symptoms experimentally contaminated honey. The set primers was designed based on published 16S rRNA gene. Using approach could amplify pathogen DNA obtain great sensitivity notable specificity. limit for spore suspension 10−2 dilution template obtained 32 spores, as determined by plate count. For artificially honey, PCR product at 10−3 170 spores. In addition, when conditions were improve specificity, able selectively no cross-reactions observed variety related species, including pulvifaciens. detection confirm presence disease, method provides reliable diagnosis AFB infected few hours.