Table 2.

Summary of PDCoV-specific RT-PCR assays.

RT-PCR Method	Target Gene	Primer or Probe	Sequences (5′-3′)	Limit of Detection	Reference
Standard One-step RT-PCR	M	primer 67F	ATCCTCCAAGGAGGCTATGC	Not defined	Wang et al. (2014a)
		primer 560R	GCGAATTCTGGATCGTTGTT
	N	primer 41F	TTTCAGGTGCTCAAAGCTCA	Not defined	Wang et al. (2014a)
		primer 735R	GCGAAAAGCATTTCCTGAAC
Standard Nested RT-PCR	N	Outer primer F	TGCTACCTCTCCGATTCCCA	Not defined	Song et al. (2015)
		Outer primer R	ATCCTGTTTGTCTGCTGGCA
		Inner primer F	GACACTGAGAAGACGGGTATGG
		Inner primer R	TAGTTGGTTTGGTAGGTGGCTC
Real-time singleplex one-step RT-PCR	M	Forward primer	ATCGACCACATGGCTCCAA	2 RNA copies per reaction	Marthaler et al. (2014b)
		Reverse primer	CAGCTCTTGCCCATGTAGCTT
		Probe	FAM-CACACCAGTCGTTAAGCAT GGCAAGCT-BHQ
	M	PDCoV-F	CGACCACATGGCTCCAATTC	0.056 TCID50per reaction	Chen et al. (2015b), Zhang et al. (2016)
		PDCoV-R	CAGCTCTTGCCCATGTAGCTT
		PDCoV-P	FAM-CACACCAGT/ZEN/CGTTAA GCATGGCAAGC-IABkFQ
	N	Forward primer	CGCTTAACTCCGCCATCAA	Not defined	Ma et al. (2015)
		Reverse primer	TCTGGTGTAACGCAGCCAGTA
		Probe	6FAM-CCCGTTGAAAACC-MGB
Real-time duplex PEDV/PDCoV RT-PCR	M	PEDV rF	GGTTGTGGCGCAGGACA	7 RNA copies per reaction for PEDV; 14 RNA copies per reaction for PDCoV	Zhang et al. (2016)
		PEDV rR	CGGCCCATCACAGAAGTAGT
		PEDV rP	FAM-CATTCTTGG/ZEN/TGGTCT TTCAATCCTGA-IABkFQ
	M	PDCoV rF	TGAGAGTAGACTCCTTGCAGGGA
		PDCoV rR	GAGAATTGGAGCCATGTGGT
		PDCoV rP	NED-TGTACCCATTGGATCCATAA-MGB
RT-iiPCR	M	PDCoV iiF	GAGAGTAGACTCCTTGCAGGGATTAT	9 RNA copies per reaction	Zhang et al. (2016)
		PDCoV iiR	GCTTGCCATGCTTAACGACTG
		PDCoV iiP	FAM-AATGCACCTCCATGTACC-MGB