Table 1.

Properties of primers used for RSV and influenza virus multiplex PCR

Amplification steps and primeraSequence (5′→3′)Gene positionMelting temp (°C)G+C content (%)Optimal annealing temp (°C)Maximum annealing temp (°C)Amplicon size (bp)Sensitivity (PFU)
Primary
 AHI ACAGATGCAGACACAATATGTHA554052631,015NAb
 AHI FIIAAACCGGCAATGGCTCCAAAHA7250
 AH3 ACAGATTGAAGTGACTAATGCHA55405262883NA
 AH3 DIIGTTTCTCTGGTACATTCCGCHA6250
 BHA AGTGACTGGTGTGATACCACTHA56505364900NA
 BHA DIITGTTTTCACCCATATTGGGCHA6545
 RSVAB FGTCTTACAGCCGTGATTAGGN63505266838NA
 RSVAB RGGGCTTTCTTTGGTTACTTCP6445
Secondary
 AHI BATAGGCTACCATGCGAACAAHA63455262944<1
 AHI EIICTTAGTCCTGTAACCATCCTHA5545
 AH3 BAGCAAAGCTTTCAGCAACTGHA63455469591<1
 AH3 CIIGCTTCCATTTGGAGTGATGCHA6550
 BHA BCATTTTGCAAATCTCAAAGCHA61355467767<1
 BHA CIITGGAGGCAATCTGCTTCACCHA6855
 RSVA FGATGTTACGGTGGGGAGTCTN64454851334<1
 RSVA RGTACACTGTAGTTAATCACAN5135
 RSVB FAATGCTAAGATGGGGAGTTCN64455061183<1
 RSVB RGAAATTGAGTTAATGACAGCN5835
  • a F, forward; R, reverse.

  • b NA, not applicable.