两种流感病毒检测方法敏感性研究

Discussion on susceptibility of two detection methods of influenza virus

目的利用实时荧光PCR相对定量方法和细胞培养法比较不同方法检测流感病毒敏感性差异。方法将流感病毒咽拭子分别接种24代和42代狗肾传代细胞（MDCK），比较不同代数MDCK细胞培养获得的毒株的血凝滴度及细胞病变（CPE）。对流感病毒咽拭子和毒株分别进行实时荧光PCR，比较实验结果CP值。结果实时荧光PCR相对定量结果显示：毒株病毒载量是咽拭子病毒载量的10^4倍～10^5倍；接种24代MDCK细胞获得毒株血凝滴度略高于接种42代MDCK细胞获得的毒株，42代MDCK细胞CPE更为明显。结论结合实时荧光PCR检测和流感病毒培养可提高流感病毒毒株分离率，24～42代MDCK传代细胞对2012年甲3型流感病毒均具有较好的敏感性，低代数MDCK传代细胞更有利于提高甲3型流感病毒分离。

Objective To compare the difference of the sensitivity of real - time PCR assay and cell culture method in detection of influenza virus. Methods The influenza virus throat swabs were inoculated to the 24th - generation and the 42nd - genera- tion of MDCK passage cells to compare the hemagglutination titer and the eytopathy （CPE）. The influenza virus throat swabs and the virus strains were detected by real - time PCR to compare the experimental results of CP. Results Real - time PCR re- sults displayed that the virus strain viral load was 10~ ~ 105 times of throat swab viral load;the strain hemagglutination titer from the 24th - generation was slightly higher than that from the 42nd - generation of MDCK cells ; the cytopathy of the 42 generation was more obviously than the 24 generation of MDCK cells. Conclusion It can increase the separation rate of virus strains by combining the real -time PCR assay and cell culture,24 -42 generation MDCK passage cells are sensitive to the influenza virus A3 in 2012, low algebraic MDCK passage cells is more propitious to improve the separation of influenza A3.