TABLE 6.
Primers and probe sequences, amplification efficiencies, and limits of detection for all PCR assays used in this study
| Type of PCR and primers and probes utilized | Sequence | Amplicon length (bp) | Amplification efficiency (%) | Limit of detection |
|---|---|---|---|---|
| enFeLV provirus LTR real-time qPCR | 137 | 95 | 10 copies/cell | |
| Forward primer | GTCTTATCCTAAGTCCACCGTTTA | |||
| Reverse primer | CTAGGCTCATCTCTAGGGTCTATC | |||
| Probe | CCTGGCCCTAAGATGGGAATGGAAA | |||
| exFeLV provirus and viremia real-time qPCR | 67 | 95 | 10 proviral copies/million cells; 200 copies/reaction; 100 copies/ml of plasma | |
| Forward primer | AGTTCGACCTTCCGCCTCAT | |||
| Reverse primer | AGAAAGCGCGCGTACAGAAG | |||
| Probe | TAAACTAACCAATCCCCATGCCTCTCGC | |||
| Feline CCR5 real-time qPCR | 115 | 95 | 10 copies/million cells | |
| Forward primer | ACGTCTACCTGCTCAACCTGG | |||
| Reverse primer | ACCGTCTTACACATCCCATCCC | |||
| Probe | TCCGACCTGCTCTTCCTCTTCACCCTCC | |||
| FeLV-B screening conventional PCR | 1,932 | NA | NA | |
| Forward primer | CAGATCAGGAACCATTCCCAGG | |||
| Reverse primer | CCTCTAACTTCCTTGTATCTCATGG | |||
| FFV real-time qPCR | 144 | 97 | 100 copies/million cells | |
| Forward primer | GGACGATCTCAACAAGGTCAACTAAA | |||
| Reverse primer | TCCACGAGGAGGTTGCGA | |||
| Probe | AGACCCCCTAGACAACAACAGCAACACT | |||
| FcaGHV-1 real-time qPCR | 113 | 97 | 10 copies/million cells | |
| Forward primer | ACATCTTCACTGGACAACTGG | |||
| Reverse primer | GTGCATTTGATGTCCTGACTG | |||
| Probe | TGAACAGCTGAGTCTCTACAAGTCTCCA | |||
| FCoV real-time qPCR | 171 | 95 | NA | |
| Forward primer | AGCAACTACTGCCACRGGAT | |||
| Reverse primer | GGAAGGTTCATCTCCCCAGT | |||
| Probe | AATGGCCACACAGGGACAACGC |