Carp edema virus surveillance in the koi trade: early detection through shipping environment sampling and longitudinal monitoring of CEV outbreaks in a wholesaler facility

Table 1 PCR primers utilised in this study for CEV detection and Sanger sequencing

PCR type		Primer name	Sequence (5’–3’)	Use	References
qPCR		CEV_TiHo_qF	CAT TTC CTA GTT TGT ATG GCA AG	Detection and quantification of CEV DNA in all samples	[25]
qPCR		CEV_TiHo_qR	TGA TGA TTG GAA TAA GAT GTC TGT C	Detection and quantification of CEV DNA in all samples	[25]
Nested PCR	First round	CEV_for_B	ATG GAG TAT CCA AAG TAC TTA G	Amplicon production for Sanger sequencing of 2019 and February 2020 samples. Second round was performed only if first round amplification was weak	[15]
	First round	CEV_rev_J	CTC TTC ACT ATT GTG ACT TTG
	Second round	CEV_for_B_int	GTT ATC AAT GAA ATT TGT GTA TTG
	Second round	CEV_rev_J_int	TAG CAA AGT ACT ACC TCA TCC
Semi-nested PCR (used with CEV_rev_J & CEV_rev_J_int)		CEV_P4a_3F	CAA CTG ACA ATG TAT CTC CAC C	Amplicon production for Sanger sequencing of batch 2022-A2 shipping water extract	Original
Conventional PCR		oPVP857	GTA CTT TAT TTG CTG CAG GAT	Amplicon production for Sanger sequencing of batch 2019-F1 shipping water extract and remaining 2020 and 2022 samples	[49]
Conventional PCR		oPVP824	GTG GTA ACT TTA CTT GTC CTC C		[49]

ISSN: 1297-9716