Detection of enteroviruses and hepatitis a virus in water by consensus primer multiplex RT-PCR

Table 1 Nucleotide sequences and positions of primers

Primers	Sequence (5’-3’)	Position	band sizes (bp)
CE (+)	ATTGGATTGGCCATCCGGTG	620-639
P (-)1	TCCACCACCACCCTCTCGACTC	1169-1190	571
P (-)2	GCATTACACTGTACGTGCAC	1271-1290	671
C (-)1	CACCAACACCCATACCTGCA	1470-1489	870
C (-)2	GCGCACATTGGCGCAATTGTG	1683-1703	1084
E (-)1	CGCACCCCATCTCAGCTTCA	1330-1349	730
E (-)2	TTCATTGTCGGCAAACCTTGA	1727-1747	1128
H (-)	CCAATCTTCCTGATCCAAAGC	1557-1577
H (+)1	AACCCTACACCTTTCCAACA	1146-1165	432
H (+)2	GATTCATTCTGCAGATTGGC	989-1008	589

+; upstream primer, -; downsream primer, 1: semi-nested primer; 2: the first PCR primer, CE; general primer, P: primers of Polioviruses; C: primers of Coxsackie viruses; E: primers of Echoviruses; H: primers of HAV

Table 2 Sensitivity of semi-nested-PCR for different viruses

Viruses	Virus concentration (PFU)
	103	102	101	100	10-1
Poliovirus	+	+	+	+ (2.4)	-
Coxsackie virus B3	+	+	+	+ (2.1)	-
Echovirus 9	+	+	+	+ (6.0)	-
Three viruses	+	+	+ (10.5)	-	-