(별표 10) 방류수산생물 검사대상 전염병의 PCR 진단 조건

병명	구분		PCR primers (5‘→3’)	PCR condition	Cycles	Product size(bp)
KHV	Protocol 1	TKF	GGGTTACCTGTACGAG	94°C for 5min 94°C for 1min, 55°C for 1min, 72°C for 1min 72°C for 10min	1 40 1	409
		TKR	CACCCAGTAGATTATGC
	Protocol 2	F	GACACCACATCTGCAAGGAG	94°C for 30s 94°C for 30s, 63°C for 30s, 72°C for 30s 72°C for 10min	1 40 1	292
		R	GACACATGTTACAATGGTCGC
RSIV	Protocol 1	1F	CTCAAACACTCTGGCTCATC	94°C for 30s, 58°C for 1min, 72°C for 1min 72°C for 5min	1 30	570
		1R	GCACCAACACATCTCCTATC
	Protocol 2	4F	CGGGGGCAATGACGACTACA	동일		568
		4R	CCGCCTGTGCCTTTTCTGGA
SVC	RT	R2	AGATGGTATGGACCCCAATACATHACNCAY	37°C for 1hr	1	714
	First	F1	TCTTGGAGCCAAATAGCTCARRTC	95°C for 1min, 55°C for 1min, 72°C for 1min 72°C for 10min	35 1
		R2	AGATGGTATGGACCCCAATACATHACNCAY
	Nested	F1	TCTTGGAGCCAAATAGCTCARRTC	동일		606
		R4	CTGGGGTTTCCNCCTCAAAGYTGY
VHS	RT(VNF)		ATGGAAGGAGGAATTCGTGAAGCG	42°C for 30min, 94°C for 5min, 4°C for 3min	1	505
	VNF		ATGGAAGGAGGAATTCGTGAAGCG	94°C for 2min 94°C for 30s, 55°C for 30s, 68°C for 60s 68°C for 7min	1 35 1
	VNR		GCGGTGAAGTGCTGCAGTTCCC
EUS	Protocol 1	Ainvad-2F	TCATTGTGAGTGAAACGGTG	95°C for 2 min 95°C for 30 s, 56°C for 45 s, 72°C for 2.5 min 72°C for 5 min	1 35 1	234
		Ainvad-ITSR1
	Protocol 2	ITS11	GGCTAAGGTTTCAGTATGTAG
		ITS23
TS	9992Forward		GCCGAAGTTTCGCAAGAAAC-	94°C for 5 min 94°C for 30 s, 65°C for 30 s, 72°C for 1 min 72°C for 5 min	1 35 1	550
	9195Reverse		CGTATAGACACAAGCACACCA
PM	Protocol 1	85	AAGTAGACAGCCGCGCTT	94°C for 1 min 94°C for 45 s, 60°C for 45 s, 60°C for 45 s 60°C for 7 min	1 40 1	231
		750	TCAATGAGAGCTTGGTCC
	Protocol 2	70F	CCTTTGYTWGAGWGTTGCCAGATG	95°C for 4min 94°C for 1min, 57°C for 1min, 65°C for 3min 65°C for 10min	1 40 1	509
		600R	CGAGTTTGCGAGTACCTGKAGAG
WSD	First	146F1	ACTACTAACTTCAGCCTATCTAG	94°C for 4min, 55°C for 1min, 72°C for 2min 94°C for 1min, 55°C for 1min, 72°C for 2min 72°C for 5min	1 39 1	1447
		146R1	TAATGCGGGTGTAATGTTCTTACGA
	Nested	146F2	GTAACTGCCCCTTCCATCTCCA	동일		941
		146R2	TACGGCAGCTGCTGCACCTTGT
	Decapod specific	143F	TGCCTTATCAGCTNTCGATTGTAG	동일		848
		143R	TTCAGNTTTGCAACCATACTTCCC