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. CONCLUSION: The TaqMan Real-time PCR assay is specific, sensitive and easy to perform.
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. CONCLUSION: The TaqMan Real-time PCR assay is specific, sensitive and easy to perform.