H5N1 type poultry grippal virus fluorescent augmentation detection kit and detection method
An avian influenza virus, detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of low separation rate, high requirements, restrictions, etc., and achieve simple and convenient sampling, high sensitivity, and judgment. Convenient and accurate results
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[0067] Utilize the TaqMan-MGB fluorescent quantitative RT-PCR method described in the present invention to detect 1 part of the lower respiratory tract aspirate specimen of the suspected H5N1 highly pathogenic avian influenza virus infection patient that occurred in Zhejiang Province in February 2006, and the result was positive (the The sample was re-examined by the Chinese Center for Disease Control and Prevention and confirmed positive for H5N1).
[0068] (1) Determination of standard curve:
[0069] The reaction system is as follows:
[0070] 5×RT-PCR buffer 5μL
[0071] Dimercaptothreitol 5mM
[0072] RNase inhibitor RNasin 0.1 unit / μL
[0073] Specific amplification upstream primer 0.48μM
[0074] Specific amplification downstream primer 0.48μM
[0075] Specific probe 0.64μM
[0076] The deoxynucleoside triphosphate mixture is:
[0077] dATP, dTTP, dCTP, dGTP each 200μM
[0078] Ampli TaqR DNA Polymerase 2.5 enzyme activity units / reaction
[0079] AMV reverse tr...
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