数字RT-PCR技术检测H3N2亚型猪流感病毒方法的建立

doi:10.16431/j.cnki.1671-7236.2017.06.038

Abstract

Abstract:

A new assay for the detection of swine influenza virus (SIV) was developed with a novel nucleic acid probe——Molecular beacon in this study. The specific primers and molecular beacon probes were designed according to the conserved region of H3 and N2 genes of SIV H3N2 subtype. A digital RT-PCR assay was developed for detection of SIV H3N2 subtype. The results showed that SIV H3N2 subtype could be identified simultaneously on this microarry with high sensitivity and reproducibility,which could reach to 10⁶ dilute viruse. The conclusion was that the digital RT-PCR method could analyze quantitatively the RNA templates.On the identification of H3N2 SIV,the digital RT-PCR method was much more scientific than Real-time quantitative PCR method.

Key words: swine influenza virus; H3N2 subtype; molecular beacon probe; digital RT-PCR; Real-time quantitative PCR; sensitivity; specificity; repeatability

CLC Number:

S852.65⁺9.5

TANG Lian-fei, YU Si-yu, ZHOU Yu. Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype[J]. , 2017, 44(6): 1847-1853.

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Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype

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