Table 1.
RT-PCR and nested PCR primers used for the detection of the group A, B and C porcine rotaviruses (PRV A–C), porcine sapovirus, porcine enteric calicivirus (designed to detect sapovirus and norovirus), transmissible gastroenteritis coronavirus (TGEV) and porcine epidemic diarrhea coronavirus (PEDV) in the fecal samples from pigs with diarrhea
Target viruses | Target genesa | Primer names | Primer sequence, 5′–3′b | Region (nt) | Size (bp) | Source or reference |
---|---|---|---|---|---|---|
PSaV and PNoV | RdRp | P289c | F: GATTACTCCAAGTGGGACTCCAC | 4568–4590 | 319 | Jiang et al. (1999a) |
P290 | R: TGA CAATGTAAT ATCACCATA | 4865–4886 | ||||
PSaV | Cap | CapsidF | F: GTGATCAACCCTTTTGAAAC | 5698–5717 | 757 | Kim et al. (2006) |
CapsidR | R: AAAGCATGATGTTGTTAGGC | 6435–6454 | ||||
PECVCnF | nF: CTCGTCATAGTAGGTGTGGC | 5890–5909 | 565 | |||
PECVCnR | nR: AAAGCATGATGTTGTTAGGC | 6435–6454 | ||||
RdRp | PEC45 | F: GTGCTCTATTGCCTGGACTA | 4312–4331 | 572 | ||
PEC46 | R: TCTGTGGTGCGGTTAGCCTT | 4864–4883 | ||||
PECnF | nF: CTCGTATGCTGAGGACACAC | 4392–4411 | 380 | |||
PECnR | nR: GAGTGTCTGTTGGCTCAATG | 4752–4771 | ||||
PRV A | VP6 | F: AAAGATGCTAGGGACAAAATTG | 58–78 | 308 | Elschner et al. (2002) | |
R: TTCAGATTGTGGAGCTATTCCA | 344–365 | |||||
nF: GACAAAATTGTCGAAGGCACATTATA | 69–94 | 121 | ||||
nR: TCGGTAGATTACCAATTCCTCCAG | 166–189 | |||||
PRV B | NSP2 | F: CTATTCAGTGTGTCGTGAGAGG | 18–40 | 434 | Gouvea et al. (1991) | |
R: GCAGACAAGCTAGCCCGCTTCG | 429–451 | |||||
PRV C | VP6 | F: CTCGATGCTACTACAGAATCAG | 994–1018 | 366 | Gouvea et al. (1991) | |
R: AGCCACATAGTTCACATTTCATCC | 1339–1359 | |||||
nF: CTCGATGCTACTACAGAATCAG | 994–1018 | 328 | ||||
nR: GGGATCATCCACGTCATGCGT | 1300–1321 | |||||
TGEV | ORF1b | F: GGGTAAGTTGCTCATTAGAAATAATGG | 7968–7994 | 1006 | Kim et al. (2000) | |
Spike | R: CTTCTTCAAAGCTAGGGACTG | 920–940 | ||||
PED | N | F: AGGAACGTGACCTCAAAGACATCCC | 812–836 | 540 | Kubota et al. (1999) | |
R: CCAGGATAAGCCGGTCTAACATTG | 1328–1351 |
RdRp: RNA dependent RNA polymerase; Cap: capsid; VP6: viral protein 6; NSP2: non-structural protein 2; ORF1b: open reading frame 1b; N: nucleocapsid.
F: upstream primer for RT-PCR; R: downstream primer for RT-PCR; nF: upstream primer for nested PCR; nR: downstream primer for nested PCR.
A primer pair p289/290 designed to detect both noroviruses and sapoviruses by RT-PCR.