Table 1 Primers and probes developed in this study
System name | Target gene | Primer and probe name | Sequence (5′-3′) | Assay specificity |
|---|---|---|---|---|
Pan-fil 28S qPCR-based system | 28S rRNA | qFil-28S-F | TTGTTTGAGATTGCAGCCCA | Filariae |
qFil-28S-P | 6FAM-CAAGTACCGTGAGGGAAAGT-TAMRA | |||
qFil-28S-R | GTTTCCATCTCAGCGGTTTC | |||
All-Wol 16S qPCR-based system | 16S rRNA | all.Wol.16S.301-F | TGGAACTGAGATACGGTCCAG | Wolbachia |
all.Wol.16S.347-P | 6FAM-AATATTGGACAATGGGCGAA-TAMRA | |||
all.Wol.16S.478-R | GCACGGAGTTAGCCAGGACT | |||
Triplex TaqMan cox1 qPCR-based system | cox1 | Fil.COI.749-F | CATCCTGAGGTTTATGTTATTATTTT | D. immitis, D. repens and A.reconditum |
D.imm.COI.777-P | 6FAM-CGGTGTTTGGGATTGTTAGTG-TAMRA | |||
D.rep.COI.871-P | 6VIC-TGCTGTTTTAGGTACTTCTGTTTGAG-TAMRA | |||
A.rec.COI.866-P | Cy5-TGAATTGCTGTACTGGGAACT-BHQ3 | |||
Fil.COI.914-R | CWGTATACATATGATGRCCYCA | |||
Duplex Wol-Diro ftsZ qPCR-based system | ftsZ | WDiro.ftsZ.490-F | AAGCCATTTRGCTTYGAAGGTG | Endosymbiotic Wolbachia of D. immitis and D. repens |
WDimm.ftsZ.523-P | 6FAM-CGTATTGCAGAGCTCGGATTA-TAMRA | |||
WDrep.ftsZ.525-P | 6VIC-CATTGCAGAACTGGGACTGG-TAMRA | |||
WDiro.ftsZ.600-R | AAACAAGTTTTGRTTTGGAATAACAAT | |||
Duplex HWs cox1 qPCR-based system | cox1 | Hw.COI.723-F | TCAGCATTTGTTTTGGTTTTT | D. immitis and A. vasorum |
D.imm.COI.777-P | 6FAM-CGGTGTTTGGGATTGTTAGTG-TAMRA | |||
A.vas.COI.813-P | 6VIC-TGACTGGGAAGAAGGAGGTG-TAMRA | |||
Hw.COI.950-R | GCASTAAAATAAGYACGAGWATC |