. 2009 Apr;58(Pt 4):408–413. doi: 10.1099/jmm.0.006098-0

Table 1.

Primers used for the detection of HRSV-A and -B, HMPV, FLU-A, -B and -C, and HPIV1, -2, -3 and -4 by nested multiplex RT-PCR

Primer	Sequence (5′→3′)	Gene	ORF position (nt)	Amplicon size (bp)
HRSV/HMPV/FLU RT-PCR
HRSVAB1	ATGGAGYTGCYRATCCWCARRRCAARTGCAAT	F	1–31	738 (HRSV-A); 738 (HRSV-B)
HRSVAB2	AGGTGTWGTTACACCTGCATTRACACTRAATTC	F	737–705	738 (HRSV-A); 738 (HRSV-B)
FluAC1	GAACTCRTYCYWWATSWCAAWGRRGAAAT	NP	319–347 (A); 346–374 (C)	722 (Flu-A); 992 (Flu-B); 739 (Flu-C)
FluB1	ACAGAGATAAAGAAGAGCGTCTACAA	NP	217–242
FluABC2	ATKGCGCWYRAYAMWCTYARRTCTTCAWAIGC	NP	1040–1009 (A); 1208–1177 (B); 1084–1053 (C)
HMPV 8426*	CCYTCAGCACCAGACACACC	NP	508–527	444
HMPV 8427*	AGATTCAGGRCCCATTTCTC	NP	951–932	444
N1	CTTGGGCGTGTCTCAAAATCT	Control	10–30	1129
N2	GTCGCCACGGTTGATGAGAGCT	Control	1137–1116	1129
HRSV/HMPV/FLU nested PCR
HRSVA3	TTATACACTCAACAATRCCAAAAAWACC	F	347–374	364 (HRSV-A)
HRSVA4	AAATTCCCTGGTAATCTCTAGTAGTCTGT	F	710–682	364 (HRSV-A)
HRSVB3	ATCTTCCTAACTCTTGCTRTTAATGCATTG	F	30–59	612 (HRSV-B)
HRSVB4	GATGCGACAGCTCTGTTGATTTACTATG	F	641–614	612 (HRSV-B)
FluAB3	GATCAAGTGAKMGRRAGYMGRAAYCCAGG	NP	718–746 (A); 892–920 (B)	302 (Flu-A); 227 (Flu-B); 114 (Flu-C)
FluC3 8425†	AAATTGGAATTTGTTCCTTT	NP	950–969
FluAC4	TCTTCAWATGCARSWSMAWKGCATGCCATC	NP	1019–990 (A); 1063–1034 (C)
FluB4	CTTAATATGGAAACAGGTGTTGCCATATT	NP	1118–1090
HMPV 8422*	CACCHATAATYTTATTATGTGTHGGTG	NP	524–550	235
HMPV 8423*	GADGATGAGCCTAAHGCTTTBC	NP	737–757	235
N3	GGGGTGTTATGAGCCATATTCAACGG	Control	108–133	888
N4	AGCCGCCGTCCCGTCAAGTCAG	Control	995–974	888
HPIV RT-PCR
HPIV1:PIP1⁺	CCTTAAATTCAGATATGTAT	HN	748–767	478
HPIV1:PIP1⁻	GATAAATAATTATTGATACG	HN	1225–1206	478
HPIV1:PIP2⁺	AACAATCTGCTGCAGCATTT	HN	803–822	508
HPIV1:PIP2⁻	ATGTCAGACAATGGGCAAAT	HN	1310–1291	508
HPIV1:PIP3⁺	CTGTAAACTCAGACTTGGTA	HN	689–708	488
HPIV1:PIP3.1⁻†	TGGAATTGAGTTTAAGCC	HN	1176–1159	488
HPIV1:PIP4⁺	CTGAACGGTTGCATTCAGGT	P	Non-coding 5′	414 (A)
HPIV1:PIP4⁻	TTGCATCAAGAATGAGTCCT	P	433–452	414 (B)
CAT gene I.C.⁺	CAGTCAGTTGCTCAATGTA	CAT	76–94	528
CAT gene I.C.⁻	GACATGGAAGCCATCAC	CAT	587–603	528
HPIV nested PCR
HPIV1:PIS1.1⁺	CCGGTAATTTCTCATACCTATG	HN	780–801	317
HPIV1:PIS1.1⁻†	CTTTGGAGCGGAGTTGTTA	HN	1096–1078	317
HPIV1:PIS2.1⁺†	CCATTTACCTAAGTGATGGA	HN	847–866	202
HPIV1:PIS2.1⁻†	GCCCTGTTGTATTTGGAAG	HN	1048–1030	202
HPIV1:PIS3.1⁺†	ACTCCCAAAGTTGATGAA	HN	811–828	356
HPIV1:PIS3.1⁻*	TTTAAGCCCTTGTCAACAAC	HN	1166–1147	356
HPIV1:PIS4.1⁺†	AAAGAATTAGGTGCAACCAG	P	76–95	245
HPIV1:PIS4⁻*	GTGTCTGATCCCATAAGCAGC	P	320–300	245
CAT gene N⁺†	CTGGAGTGAATACCACGA	CAT	315–332	149
CAT gene N⁻†	TGGTGAAACTCACCCAG	CAT	463–447	149
HPIV-4 subtyping
HPIV4A:PIS4A⁺	ATGATGGTGGAACCAAGATT	P	146–165	175 (A); 82 (B)
HPIV4B:PIS4B⁺	AACCAGGGAAACAGAGCTC	P	239–257
HPIV4:PIS4⁻*	GTGTCTGATCCCATAAGCAGC	P	320–300

*Newly created primers.

†Modified from the original primers.