Table 1.

Oligonucleotides and PCR conditions used for detection of canine viral pathogens

Virus	Test	Reference	Oligonucleotides	PCR conditionsa	Amplicon size or fluorescence signal
CAV-1/CAV-2	PCR	Hu et al. (2001)	HA1 (+), CGCGCTGAACATTACTACCTTGTC	94 °C for 30 s	CAV-1, 508 bp
			HA2 (−), CCTAGAGCACTTCGTGTCCGCTT	58 °C for 1 min	CAV-2, 1030 bp
				72 °C for 1 min
MRV	RT-PCR	Leary et al. (2002)	L1-rv5 (+), GCATCCATTGTAAATGACGAGTCTG	94 °C for 30 s	416 bp
			L1-rv6 (−), CTTGAGATTAGCTCTAGCATCTTCTG	50 °C for 30 s
				72 °C for 30 s
MRV-1	RT-PCR	Decaro et al. (2005a)	S1-R1F (+), GGAGCTCGACACAGCAAATA	94 °C for 30 s	505 bp
			S1-R1R (−), GATGATTGACCCCTTGTGC	53 °C for 30 s
				72 °C for 30 s
MRV-2	RT-PCR	Decaro et al. (2005a)	S1-R2F (+), CTCCCGTCACGGTTAATTTG	94 °C for 30 s	394 bp
			S1-R2R (−), GATGAGTCGCCACTGTGC	53 °C for 30 s
				72 °C for 30 s
MRV-3	RT-PCR	Decaro et al. (2005a)	S1-R3F (+), TGGGACAACTTGAGACAGGA	94 °C for 30 s	326 bp
			S1-R3R (−), CTGAAGTCCACCRTTTTGWA	53 °C for 30 s
				72 °C for 30 s
RV	RT-PCR	Gouvea et al. (1994)	Beg9 (+), GGCTTTAAAAGAGAGAATTTCCGTCTGG	94 °C for 1 min	1060 bp
			End9 (−), GGTCACATCATACAATTCTAATCTAAG	55 °C for 2 min
				72 °C for 1 min
CaCV	RT-PCR	Jiang et al. (1999)	289 (+), TGACAATGTAATCATCACCATA	94 °C for 1 min	300–320 bp
			290 (−), GATTACTCCAAGTGGGACTCCAC	48 °C for 1 min
				72 °C for 1 min
FCV	RT-PCR	Marsilio et al. (2005)	Cali 1 (+), AACCTGCGCTAACGTGCTTA	94 °C for 1 min	924 bp
			Cali 2 (−), CAGTGACAATACACCCAGAAG	57 °C for 45 s
				72 °C for 1 min
CaHV-1	PCR	Schulze and Baumgartner (1998)	CHV1 (+), TGCCGCTTTTATATAGATG	94 °C for 1 min	493 bp
			CHV2 (+), AAGCTGTGTAAAAGTTCGT	53 °C for 1 min
				72 °C for 1 min
CPV-2 (all types)	Real-time PCRa	Decaro et al. (2005c)	CPV-For (+), AAACAGGAATTAACTATACTAATATATTTA	94 °C for 15 s	FAM
			CPV-Rev (−), AAATTTGACCATTTGGATAAACT	52 °C for 30 s
			CPV-Pb (+), FAM–TGGTCCTTTAACTGCATTAAATAATGTACC–TAMRA	60 °C for 1 min
CPV-2 (types 2a/2b)	Real-time PCRb,c	Decaro et al. (2006)	CPVa/b-For (+), AGGAAGATATCCAGAAGGAGATTGGA	94 °C for 15 s	CPV-2a, VIC
			CPVa/b-Rev (−), CCAATTGGATCTGTTGGTAGCAATACA	60 °C for 1 min	CPV-2b, FAM
			CPVa-Pb (+), VIC–CTTCCTGTAACAAATGATA–MGB
			CPVb1-Pb (+), FAM–CTTCCTGTAACAGATGATA–MGB
CPV-2 (types 2b/2c)	Real-time PCRb,c	Decaro et al. (2006)	CPVb/c-For (+), GAAGATATCCAGAAGGAGATTGGATTCA	94 °C for 15 s	CPV-2b, FAM
			CPVb/c-Rev (−), ATGCAGTTAAAGGACCATAAGTATTAAATATATTAGTATAGTTAATTC	60 °C for 1 min	CPV-2c, VIC
			CPVb2-Pb (+), FAM–CCTGTAACAGATGATAAT–MGB
			CPVc-Pb (−), VIC–CCTGTAACAGAAGATAAT–MGB
CDV	Real-time RT-PCRa	Elia et al. (2006)	CDV-F (+), AGCTAGTTTCATCTTAACTATCAAATT	94 °C for 15 s	FAM
			CDV-r (+), TTAACTCTCCAGAAAACTCATGC	48 °C for 30 s
			CDV-Pb (−), FAM–ACCCAAGAGCCGGATACATAGTTTCAATGC–TAMRA	60 °C for 1 min
CCoV	Real-time RT-PCRa	Decaro et al. (2004b)	CCoV-For (+), TTGATCGTTTTTATAACGGTTCTACAA	94 °C for 15 s	FAM
			CCoV-Rev (−), AATGGGCCATAATAGCCACATAAT	60 °C for 1 min
			CCoV-Pb (+), FAM–ACCTCAATTTAGCTGGTTCGTGTATGGCATT–TAMRA
CCoV type I	Real-time RT-PCRa	Decaro et al. (2005d)	CCoVI-F (+), CGTTAGTGCACTTGGAAGAAGCT	94 °C for 15 s	FAM
			CCoVI-R (−), ACCAGCCATTTTAAATCCTTCA	53 °C for 30 s
			CCoVI-Pb (+), FAM–CCTCTTGAAGGTACACCAA–TAMRA	60 °C for 1 min
CCoV type II	Real-time RT-PCRa	Decaro et al. (2005d)	CCoVII-F (+), TAGTGCATTAGGAAGAAGCT	94 °C for 15 s	FAM
			CCoVII-R (−), AGCAATTTTGAACCCTTC	48 °C for 30 s
			CCoVII-Pb (+), FAM–CCTCTTGAAGGTGTGCC–TAMRA	60 °C for 1 min

CAV, canine adenovirus; MRV, mammalian reovirus; RV, rotavirus, CaCV, canine calicivirus; FCV, feline calicivirus; CaHV, canid herpesvirus; CPV, canine parvovirus; CDV, canine distemper virus; CCoV, canine coronavirus; +, sense; −, antisense.

^aPCR conditions are referred to 40 cycles of amplification consisting of denaturation, primer annealing and extension.

^bReal-time PCR with conventional TaqMan probes.

^cReal-time PCR with minor groove binder probes.