[Design and rapid evaluation of a TaqMan assay for the detection of influenza A viruses].
OBJECTIVE
To design and rapidly evaluate a TaqMan assay for detecting influenza A viruses.


METHODS
The probe and the primers of the assay were designed with the software packages of DNA Star and Primer Premier 5.0. Their specificity and conservation were verified through Blast in GenBank and electronic hybridization. The assay's sensitivity was compared with the standard RT-PCR.


RESULTS
The designed primers and probe were confirmed to be very specific and conserved. The assay was 3-27 folds more sensitive than the standard RT-PCR. The RT and PCR steps could be simplified into one step.


CONCLUSIONS
The TaqMan Real-time PCR assay is specific, sensitive and easy to perform.
Chen
Ji-ming
JM
National Diagnostic Center of Animal Exotic Infectious Diseases, Animal Quarantine Institute, Ministry of Agriculture, Qingdao, 266032, China.
Wang
Zhi-liang
ZL
Pang
Zeng-chang
ZC
Sun
Cheng-ying
CY
Sun
Ying-xue
YX
Song
Cui-ping
CP
Yi
Ying
Y
D000818:Animals; D001717:Birds; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity