A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon with in SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.

• Ji-Young Choi(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
• Jong-Gill Kim(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
• Young-Cheol Choi(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
• Won-Tae Kim(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
• Gil-Sang Jeong(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
• Seok-Jo Hwang(Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)