A PCR DETECTION METHOD FOR RAPID IDENTIFICATION OF MELISSOCOCCUS PLUTON IN HONEYBEE LARVAE

Melissococcus pluton is the causative agent of European foulbrood, a d isease of honeybee larvae. This bacterium is particularly difficult to isolate because of its stringent growth requirements and competition from other bacteria. PCR was used selectively to amplify specific rRNA gene sequences of M. pluton from pure culture, from crude cell lysate s, and directly from infected bee larvae. The PCR primers were designe d from M. pluton 16S rRNA sequence data. The PCR products were visuali zed by agarose gel electrophoresis and confirmed as originating from M . pluton by sequencing in both directions. Detection was highly specif ic, and the probes did not hybridize with DNA from other bacterial spe cies tested. This method enabled the rapid and specific detection and identification of M. pluton from pure cultures and infected bee larvae .