126.96.36.199.2. Multiplex PCR for detection of Nosema and differentiation between N. apis, N. bombi and N. ceranae
For multiplex PCR amplification of partial 16S rRNA (=SSU rRNA) gene fragments, we recommend the following primer combination, though others from Table 1 (standard or qPCR primers) may be more suitable for different purposes and in different laboratories:
designed based on alignment of all available sequence data in GenBank of the
16S rRNA gene from N. apis, N. bombi and N. ceranae.
Mnapis-F forward primer: 5’-GCATGTCTTTGACGTACTATG-3’
Mnbombi-F forward primer: 5’-TTTATTTTATGTRYACMGCAG-3’
Mnceranae-F forward primer: 5’- CGTTAAAGTGTAGATAAGATGTT-3’
Muniv-R: reverse primer: 5’-
Note that the Mnbombi-F primer contains variable sites to account for the sequence diversity observed for this species.
PCR product size:
- for N. ceranae: 143 bp
- for N. bombi: 171 bp
- for N. apis: 224 bp