tetano
Editor, Senior Moderator
J Virol Methods. 2015 Mar 5. pii: S0166-0934(15)00065-8. doi: 10.1016/j.jviromet.2015.02.025. [Epub ahead of print]
[h=1]Development of a TaqMan MGB RT-PCR for the rapid detection of H3 subtype avian influenza virus circulating in China.[/h] Teng Q[SUP]1[/SUP], Shen W[SUP]2[/SUP], Yan D[SUP]2[/SUP], Yan L[SUP]1[/SUP], Li X[SUP]1[/SUP], Li G[SUP]1[/SUP], Yang J[SUP]1[/SUP], Li Z[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Previous studies demonstrated that the H3 avian influenza virus (AIV) in China is isolated most frequently from wild birds and live poultry markets. However, there is no subtype-specific real-time polymerase chain reaction (RT-PCR) available for the rapid and highly sensitive identification of H3 AIV. In this study, a TaqMan minor groove binder (MGB) probe and a pair of primers were designed based on a conserved region in the hemagglutinin gene of H3 AIV. These were used to generate an H3-MGB RT-PCR assay that recognizes only H3 AIV. The detection limit of the H3-MGB RT-PCR was 10 copies of DNA per reaction when 10-fold serial dilutions of T-H3HA plasmid were used as the template. This was 1000-times more sensitive than conventional RT-PCR. In experimental samples obtained from oropharyngeal swabs or cloacal swabs, the virus was detected in all ducks using H3-MGB RT-PCR, whereas only one duck tested positive for the virus in oropharyngeal swabs tested using conventional RT-PCR. The H3-MGB RT-PCR assay developed in this study is a sensitive and rapid tool for screening H3 AIV in China.
Copyright ? 2015. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Detection; H3 subtype; Minor groove binder; sensitivity
PMID: 25745957 [PubMed - as supplied by publisher]
[h=1]Development of a TaqMan MGB RT-PCR for the rapid detection of H3 subtype avian influenza virus circulating in China.[/h] Teng Q[SUP]1[/SUP], Shen W[SUP]2[/SUP], Yan D[SUP]2[/SUP], Yan L[SUP]1[/SUP], Li X[SUP]1[/SUP], Li G[SUP]1[/SUP], Yang J[SUP]1[/SUP], Li Z[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Previous studies demonstrated that the H3 avian influenza virus (AIV) in China is isolated most frequently from wild birds and live poultry markets. However, there is no subtype-specific real-time polymerase chain reaction (RT-PCR) available for the rapid and highly sensitive identification of H3 AIV. In this study, a TaqMan minor groove binder (MGB) probe and a pair of primers were designed based on a conserved region in the hemagglutinin gene of H3 AIV. These were used to generate an H3-MGB RT-PCR assay that recognizes only H3 AIV. The detection limit of the H3-MGB RT-PCR was 10 copies of DNA per reaction when 10-fold serial dilutions of T-H3HA plasmid were used as the template. This was 1000-times more sensitive than conventional RT-PCR. In experimental samples obtained from oropharyngeal swabs or cloacal swabs, the virus was detected in all ducks using H3-MGB RT-PCR, whereas only one duck tested positive for the virus in oropharyngeal swabs tested using conventional RT-PCR. The H3-MGB RT-PCR assay developed in this study is a sensitive and rapid tool for screening H3 AIV in China.
Copyright ? 2015. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Detection; H3 subtype; Minor groove binder; sensitivity
PMID: 25745957 [PubMed - as supplied by publisher]