A real-time PCR assay for the monitoring of influenza A virus in wild birds.
A screening system including a new real-time PCR assay for the monitoring of influenza A virus in wild birds was developed. The real-time PCR assay uses SYBR green chemistry and the primers are targeting the matrix gene of influenza A virus. The performance of the assay was compared with two other assays, one assay also using SYBR green chemistry and one assay using TaqMan chemistry, i.e. a specific probe. A total of 45 fecal bird samples were analysed for influenza A virus in three different PCR reactions. Overall, 26 samples were positive in at least one of the three real-time PCR assays. Of the 26 samples, 18 were positive by all three reactions. Eight samples were found positive exclusively by the two SYBR green reactions, six of which were detected by both SYBR green reactions. Of the 26 positive samples, 15 samples were verified as positive either by virus isolation or influenza A M2-gene PCR. The results showed that the two SYBR green systems had a higher performance regarding the detection of influenza A as compared to the PCR reaction using a specific probe.
Karlsson
Malin
M
Swedish Institute for Infectious Disease Control, SE-171 82 Stockholm, Sweden.
Wallensten
Anders
A
Lundkvist
Ake
A
Olsen
Björn
B
Brytting
Maria
M
D000818:Animals; D000835:Animals, Wild / Q000821:virology*; D004372:Ducks / Q000821:virology*; D005243:Feces / Q000821:virology; D005456:Fluorescent Dyes; D009980:Influenza A virus / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D012367:RNA, Viral