不同抽提方式在流行性感冒病毒检测中的抽提效率研究  

Extraction efficiency of different extraction methods in detection of influenza viruses

作　　者：张相猛 周志豪 潘俊锋[1] 徐秋芳[1] ZHANG Xiangmeng;ZHOU Zhihao;PAN Junfeng;XU Qiufang(Department of Microbiological Testing,Qingpu District Center for Disease Control and Prevention,Shanghai 201700,China)

机构地区：[1]上海市青浦区疾病预防控制中心微生物检验科,上海201700

出　　处：《检验医学与临床》2023年第13期1900-1902,1906,共4页Laboratory Medicine and Clinic

基　　金：上海市青浦区卫生健康委员会科研课题(W2021-43)。

摘　　要：目的比较普通抽提法和快速抽提法对实时荧光定量聚合酶链反应(PCR)检测流行性感冒(流感)病毒结果的影响。方法采用同一品牌快速核酸抽提试剂和普通核酸抽提试剂抽提经稀释不同倍数的流感病毒毒株,采用实时荧光定量PCR测定抽提核酸的Ct值,比较两种方法的抽提效率和重复性。结果普通抽提法在不同稀释倍数下Ct值均值均小于快速抽提法;普通抽提法在原液至107稀释倍数下Ct值变异系数(CV)均小于快速抽提法;原液至106稀释倍数,普通抽提法和快速抽提法检出率均为100%;107稀释倍数下,普通抽提法和快速抽提法检出率分别为100%、50%;108稀释倍数下,普通抽提法和快速抽提法检出率分别为75%、0;稀释至109稀释倍数后,两者检出率均为0。结论普通抽提法较快速抽提法抽提效率高、重复性好、检出率高,在抽提高病毒含量标本时两种方法均可使用,抽提低病毒含量标本或复杂标本中推荐使用普通抽提法。Objective To compare the effects between the common extraction method and rapid extraction method on the results of real-time fluorescence quantitative PCR in detecting influenza virus.Methods The same brand of rapid nucleic acid extraction reagent and common nucleic acid extraction reagent were used to extract the influenza strains diluted in different multiples.The Ct value of the extracted nucleic acid was determined by the real-time fluorescence quantitative PCR.The extraction efficiency and repeatability were compared between the two methods.Results The average Ct value under different dilution ratios in the common extraction method was smaller than that in the rapid extraction method.The coefficient of variation(CV)of Ct value under the stock solution to 107 dilution ratios in the common extraction method was smaller that in the rapid extraction method;the detection rates of stock solution to 106 dilution ratios by the common extraction method and rapid extraction method were 100%,the detection rates under the 107 dilution ratios in the common extraction method and rapid extraction method were 100%and 50%respectively,which under the 108 dilution ratios were 75%and 0 respectively,and which under 109 dilution ratios all were 0.Conclusion The common extraction method has higher extraction efficiency,better repeatability and higher detection rate than the rapid extraction method.Both methods could be used when extracting the high virus content,and the common extraction method is recommended when extracting the sample with low virus content or complex samples.

关 键 词：流行性感冒病毒 核酸抽提 实时荧光定量聚合酶链反应 

分 类 号：R446.5[医药卫生—诊断学]