3种流感病毒检测方法灵敏性比较

Sensitivity comparison of three detection methods for influenza virus

目的比较real-time PCR、RT-PCR和细胞培养法在流感病毒检测敏感性的差异,为实验室进行流感病毒相关实验筛选方法提供依据。方法将血凝滴度为1∶16甲型H1N1、H3N2、Victoria、Yamagata 4种流感病毒亚型进行2、5、101、102、103、104、105、106、107、108、109、1010稀释后,分别用real-time PCR、RT-PCR和细胞培养法检测,比较检测的结果。结果 H1N1流感病毒分别用real-time PCR、RT-PCR和细胞培养法能检测到的最低稀释浓度为1010、108、1010,H3N2流感病毒用3种检测方法能检测到的最低稀释浓度为1010、105、105,Victoria流感病毒用3种检测方法检测到的最低稀释浓度为107、106、106,Yamagata流感病毒用3种检测方法检测的最低稀释浓度为106、102、102。结论在处理流感疫情时,选用灵敏度高、用时短的real-time PCR法,用细胞培养法进行验证。

Objective To compare the sensitivity differences of real-time quantitative polymerase chain reaction（ PCR）,reverse transcription polymerase chain reaction（ RT-PCR） and cell culture in influenza virus detection,so as to provide basis for method choosing in the laboratory experiments of influenza virus. Methods H1 N1,H3 N2,Victia,Yamagata influenza virus with hemagglutination titer of 1∶16 were diluted to 2,5,101,102,103,104,105,106,107,108,109,1010,and then detected by real-time PCR,RT-PCR and cell culture,and the detection results were compared. Results The lowest dilution concentration of influenza H1 N1 virus detected by real-time PCR,RT-PCR and cell culture method respectively were 1010,108,1010;the lowest dilution concentration detected by three detection methods in H3 N2 were 1010,105,105; the lowest dilution concentration detected by three detection methods in Victoria were 107,106,106; the lowest dilution concentration detected by three detection methods in Yamagata were 106,102,102. Conclusion When dealing with influenza outbreaks,we should choose real-time PCR which is the highest sensitivity and the short time in the three methods and make the detection results verified by cell culture.