Development and application of a triplex real-time PCR assay for simultaneous detection of avian influenza virus, Newcastle disease virus, and duck Tembusu virus

Table 6 Primers and Probes

Name ^a	Sequence (5′-3′) ^b	Genomic region	Target genes	Amplicon size(bp)
AIV-F	AGGGTTTGTGTTCACGCTC	180–198		186
AIV-R	CCGGTTGAGTAGCTGAGTGC	346–365	M
AIV-P	ROX - CCGTGCCCAGTGAGCGAGGAC - BHQ1	210–233
NDV-F	GACTCAACTCTTGGGCATACA	837–857		172
NDV-R	TGAGGTGTCAAGCTCTTCTAT	988–1008	F
NDV-P	FAM – CAGTCGGGAACCTAAATAATATGCGTGC - BHQ-1	872–899
DTMUV-F	CAGAGACTGGTTTCATGA	642–659		122
DTMUV-R	AAGCCACCACTGATTGTT	746–763	E
DTMUV-P	JOE - TTACCATGGACAGGGTCATCAGC - BHQ-2	667–689

^a F: forward primer, R: reverse primer, P: TaqMan probe
^b ROX: 6-carboxy-x-rhodamine, FAM: 6-carboxy-fluorescein, BHQ-1:Black Hole Quencher 1, BHQ-2: Black Hole Quencher

ISSN: 1746-6148