论文部分内容阅读
目的对实时荧光定量PCR法和细胞分离培养法2种流感病毒检测方法和结果进行比较研究,为2种方法各自的实用领域提供参考依据。方法在流感高峰季节收集流感哨点医院流感样病例标本(ILI),同时进行实时荧光定量PCR检测和狗肾传代细胞培养,比较2种方法检测结果的差异。结果 1 025份ILI病例样本中,实时荧光定量PCR、细胞培养检测流感病毒核酸阳性率分别为26.63%、12.58%,差异有统计学意义(χ2=64.165,P<0.05)。273份流感核酸阳性ILI病例样本中,分离病毒株118株,分离率为43.22%;752份阴性样本中,分离病毒株11株,分离率为1.46%。实时荧光定量PCR阳性标本与阴性标本分离率差异有统计学意义(χ2=94.712,P<0.05)。结论实时荧光定量PCR具有高度灵敏性,检测速度快,实用于流感疫情应急检测、临床诊断、常规监测;细胞培养可获得毒株,用来开展抗原性、基因特性和耐药性等深入研究。
Objective To compare the detection methods and results of two kinds of influenza viruses by real-time fluorescence quantitative PCR and cell isolation and culture methods, and provide reference for their respective practical fields. Methods Influenza sentinel hospital influenza-like specimens (ILI) were collected during the peak season of influenza. Real-time fluorescent quantitative PCR and dog kidney passaged cell culture were also performed. The differences between the two methods were compared. Results The positive rate of influenza virus nucleic acid detected by real-time fluorescence quantitative PCR and cell culture in 1 025 ILI cases were 26.63% and 12.58%, respectively, with significant difference (χ2 = 64.165, P <0.05). Among 273 samples of ILI positive for influenza nucleic acid, 118 isolates were isolated with the isolation rate of 43.22%. Among 752 negative samples, 11 strains were isolated and the isolation rate was 1.46%. There was a significant difference in the separation rate between the positive and negative specimens of real-time PCR (χ2 = 94.712, P <0.05). Conclusion Real-time PCR is highly sensitive and rapid in detection. It is practically used in emergency detection, clinical diagnosis and routine monitoring of influenza epidemic. Strain can be obtained by cell culture for further research on antigenicity, gene characteristics and drug resistance.