Table 1.

 PCR reactions used for detection of respiratory viruses

Viruses detected	Methods	Round	Sequences of primers	Target genes	Thermal profile	Size of PCR products	References
IFVs  A, B, and C; RSVs  A and B	Multiplex nested RT‐PCR	1st	FluAC1: 5′‐GAACTCRTYCYWWATSWCAAWGRRGAAAT‐3′	NP	48°C for 45 min; 94°C for 3 min; 94°C for 30 s, 55°C for 60 s, 72°C for 60 s, 45 cycles. 72°C for 10 min	IFV  A: 301 bp IFV  B: 226 bp IFV  C: 111 bp RSV  A: 363 bp RSV  B: 611 bp	[1]
			FluB1: 5′‐ACAGAGATAAAGAAGAGCGTCTACAA‐3′
			FluABC2: 5′‐ATKGCGCWYRAYAMWCTYARRTCTTCAWAIGC‐3′
			RSVAB1: 5′‐ATGGAGYTGCYRATCCWCARRRCAARTGCAAT‐3′	F
			RSVAB2: 5′‐AGGTGTWGTTACACCTGCATTRACACTRAATTC‐3′
		2nd	FluAB3: 5′‐GATCAAGTGAKMGRRAGYMGRAAYCCAGG‐3′	NP	94°C for 3 min; 94°C for 30 s, 55°C for 60 s, 72°C for 45 s, 35 cycles; 72°C for 10 min
			FluC3: 5′‐AAATTGGAATTTGTTCCTTTCAAGGGACA‐3′
			FluAC4: 5′‐TCTTCAWATGCARSWSMAWKGCATGCCATC‐3′
			FluB4: 5′‐CTTAATATGGAAACAGGTGTTGCCATATT‐3′
			RSVA3: 5′‐TTATACACTCAACAATRCCAAAAAWACC‐3′	F
			RSVA4: 5′‐AAATTCCCTGGTAATCTCTAGTAGTCTGT‐3′
			RSVB3: 5′‐ATCTTCCTAACTCTTGCTRTTAATGCATTG‐3′
			RSVB4: 5′‐GATGCGACAGCTCTGTTGATTTACTATG‐3′
PIVs  1–4; HRV; EV	Multiplex nested RT‐PCR	1st	1PIV13: 5′‐AGGWTGYSMRGATATAGGRAARTCATA‐3′	HA	48°C for 45 min; 94°C for 3 min; 94°C for 30 s, 55°C for 60 s, 72°C for 60 s, 45 cycles; 72°C for 10 min	PIV  1: 439 bp PIV  2: 297 bp PIV  3: 390 bp PIV  4: 174 bp EV: 226 bp (200–232 bp) RV: 110 bp (100–120 bp)	[19]
			2PIV13: 5′‐CTWGTATATATRTAGATCTTKTTRCCTAGT‐3′
			1PIV2: 5′‐TAATTCCTCTTAAAATTGACAGTATCGA‐3′
			1PIV4: 5′‐ATCCAGARRGACGTCACATCAACTCAT‐3′	5′NCR‐HA
			2PIV24: 5′‐TRAGRCCMCCATAYAMRGGAAATA‐3′	HA
			1‐EV/RVF: 5′‐CTCCGGCCCCTGAATRYGGCTAA‐3′	5′NCR‐VP4/VP2
			2‐EV/RVR: 5′‐TCIGGIARYTTCCASYACCAICC‐3′
		2nd	3PIV13: 5′‐ACGACAAYAGGAARTCATGYTCT‐3′	HA	94°C for 3 min; 94°C for 30 s, 55°C for 60 s, 72°C for 30 s, 35 cycles; 72°C for 10 min
			4PIV1: 5′‐GACAACAATCTTTGGCCTATCAGATA‐3′
			4PIV3: 5′‐GAGTTGACCATCCTYCTRTCTGAAAAC‐3′
			3PIV24: 5′‐CYMAYGGRTGYAYTMGAATWCCATCATT‐3′
			4PIV2: 5′‐GCTAGATCAGTTGTGGCATAATCT‐3′
			4PIV4: 5′‐TGACTATRCTCGACYTTRAAATAAGG‐3′
			3‐EV/RVF: 5′‐ACCRASTACTTTGGGTRWCCGTG‐3′	5′NCR‐VP4/VP2
			4‐EV/RVR: 5′‐CTGTGTTGAWACYTGAGCICCCA‐3′
HCoV	RT‐PCR		HCoVFc: 5′‐GGTTGGGACTATCCTAAGTGTGA‐3′	Pol	48°C for 45 min; 94°C for 3 min; 94°C for 30 s, 48°C for 30 s, 72°C for 45 s, 40 cycles; 72°C for 10 min	440 bp	[20]
			HCoVRc: 5′‐CCATCATCAGATAGAATCATCATA‐3′
HMPV	RT‐PCR		HMPVR: 5′‐CATGCCCACTATAAAAGGTCAG‐3′	L	48°C for 45 min; 94°C for 3 min; 94°C for 30 s, 55°C for 30 s, 72°C for 45 s, 40 cycles; 72°C for 10 min	171 bp	[21]
			HMPVF: 5′‐CACCCCAGTCTTTCTTGAAA‐3′
ADV	PCR		AdvF: 5′‐GCCSCARTGGKCWTACATGCACATC‐3′	Hexon	94°C for 3 min; 94°C for 30 s, 55°C for 30 s, 72°C for 45 s, 35 cycles; 72°C for 10 min	301 bp	[22]
			AdvR: 5′‐CAGCACSCCICGRATGTCAAA‐3′

EV, enterovirus; HCoV, human coronavirus; HMPV, human metapneumovirus; HRV, human rhinovirus; IFV, influenza virus; PIV, parainfluenza virus; RSV, respiratory syncytial virus.