Table 2.
Virus | Gene | Primer sequence | PCR conditions | Primer final concentration | Reference |
---|---|---|---|---|---|
EV | 5' Noncoding region | EV1: 5'-ATTGTCACCATAAGCAGCCA-3' EV2: 5'-TCCGGCCCCTGAATGCGGCTAATCC-3' EV3: 5'-ACACGGACACCCAAAGTAGTCGGTTCC-3' EV4: 5' TCCGGCCCCTGAATGCGGCTAATCC3' |
PCRI: EV1/EV2 Activation cycle (95°C 5 min) 30 cycles: 95°C 45s, 55°C 45 s, and 70°C 45s PCRII: EV3/EV4 Activation cycle (95°C 5 min) 30 cycles: 95°C 45s, 55°C 45 s, and 70°C 45 s |
EV1: 6.1 mM EV2: 7.6 mM EV3: 8.2 mM EV3: 7.6 mM |
37 |
HAV | VP1 region | HA1: 5'-TTGCTCCTCTTTATCATGCTATG-3' HA3: 5'-TGGTTAAATCTAATGGTCCTCTATA-3' |
Activation cycle (95°C 5 min) 40 cycles: 95°C 30s, 46°C 30 s, and 70°C 30s |
HA1: 8.7 mM HA3: 9.6 mM |
38 |
RV | NSP-3 | ROTAS1: 5' ACCATCTTCACgTAACCCTC 3' ROTAS2: 5'-ACCATCTACACATGACCCTC-3' ROTAA: 5'-CACATAACGCCCCTATAGCC-3' ROTAP: [6FAM]-GGGGATGAGCACAATAGTTAAAAGCTA ACACTGTCAA -[BBQ] |
Activation cycle (95°C 5 min) 35 cycles: 95 ° 20 s, 60°C 40 s |
ROTAS1: 0.2 pM ROTAS2: 0.2 pM ROTAA: 0.2 pM ROTAAP: 0.2 pM |
39 |
NoVGI | ORF1 | NVG1F: 5'CGYTGGATGCGNTTCCATGA 3' NVG1R: 5'-GTCCTTAGACGCCATCATC-3' G1-prob: [6FAM]-AGATYGCGRTCYCCTGTCCA-[BHQ1] |
Activation cycle (95°C 5 min) 35 cycles: 95 ° 15 s, 56°C 60 s |
NVG1F: 0.2 pM NVG1R: 0.2 pM G1-prob: 0.2 pM |
40 |
NoVGII | ORF1 | NVG2F: 5'ATGTTYAGRTGGATGAGRTTYTC 3' COG2R: 5'-TCGACGCCATCTTCATTCACA-3' G2-prob: [JOE]TGGGAGGGCGATCGCAATCT[BHQ1] |
Activation cycle (95°C 5 min) 35 cycles: 95 ° 15 s, 56°C 60 s |
NVG2F: 0.2 pM COG2R: 0.2 pM G2-prob: 0.2 pM |
40 |