TABLE 1.

Properties of primers used for RSV and influenza virus multiplex PCR

Amplification steps and primera	Sequence (5′→3′)	Gene position	Melting temp (°C)	G+C content (%)	Optimal annealing temp (°C)	Maximum annealing temp (°C)	Amplicon size (bp)	Sensitivity (PFU)
Primary
AHI A	CAGATGCAGACACAATATGT	HA	55	40	52	63	1,015	NAb
AHI FII	AAACCGGCAATGGCTCCAAA	HA	72	50
AH3 A	CAGATTGAAGTGACTAATGC	HA	55	40	52	62	883	NA
AH3 DII	GTTTCTCTGGTACATTCCGC	HA	62	50
BHA A	GTGACTGGTGTGATACCACT	HA	56	50	53	64	900	NA
BHA DII	TGTTTTCACCCATATTGGGC	HA	65	45
RSVAB F	GTCTTACAGCCGTGATTAGG	N	63	50	52	66	838	NA
RSVAB R	GGGCTTTCTTTGGTTACTTC	P	64	45
Secondary
AHI B	ATAGGCTACCATGCGAACAA	HA	63	45	52	62	944	<1
AHI EII	CTTAGTCCTGTAACCATCCT	HA	55	45
AH3 B	AGCAAAGCTTTCAGCAACTG	HA	63	45	54	69	591	<1
AH3 CII	GCTTCCATTTGGAGTGATGC	HA	65	50
BHA B	CATTTTGCAAATCTCAAAGC	HA	61	35	54	67	767	<1
BHA CII	TGGAGGCAATCTGCTTCACC	HA	68	55
RSVA F	GATGTTACGGTGGGGAGTCT	N	64	45	48	51	334	<1
RSVA R	GTACACTGTAGTTAATCACA	N	51	35
RSVB F	AATGCTAAGATGGGGAGTTC	N	64	45	50	61	183	<1
RSVB R	GAAATTGAGTTAATGACAGC	N	58	35

^aF, forward; R, reverse. 

^bNA, not applicable.