流感病毒快速检测方法特异性和敏感性研究  

Research on the specificity and sensitivity of rapid detection methods of influenza virus

作　　者：尤凤兴[1] 居丽雯[2] 张敬平[1] 施强[2] 马广源[1] 吉杏生[1] 吴家林[1] 凌霞[1] 

机构地区：[1]无锡市疾病预防控制中心,江苏无锡214023 [2]复旦大学公共卫生学院,上海200032

出　　处：《检验医学》2009年第8期598-601,共4页Laboratory Medicine

基　　金：国家卫生部资助项目(WKJ2006-2-10)

摘　　要：目的比较多重逆转录聚合酶链反应（mRT—PCR）和胶体金法检测流感病毒的敏感性和特异性。方法采用mRT—PCR和胶体金法检测经细胞培养和红细胞凝集抑制试验（HI）鉴定为A／H1、A／H3、B型流感病毒的鼻咽拭标本78份，阴性鼻咽拭标本40份，并对已稀释成10^1～10^5病毒半数感染量（TCID50）／0．1mL的3株H1、H3、B型流感病毒进行敏感性试验。结果toRT—PCR对A型流感病毒的检测敏感性为44．8％，特异性为97．5％；对B型流感病毒检测敏感性为20．0％，特异性为100．0％。胶体金法对A型流感病毒检测敏感性为44．8％，特异性为100．0％；对B型流感病毒检测敏感性为5．0％，特异性为97．5％。对经20代内狗肾细胞（MDCK）分离培养阳性的4份A／H1、4份A／H3和3份B型流感培养液，mRT—PCR和胶体金法检测的阳性符合率与型别符合率均为100．0％。mRT—PCR检测流感病毒敏感性为10^3TCID50／0．1mL；胶体金法检测的敏感性为10^3TCID50／0．1mL。结论toRT—PCR和胶体金法的敏感性与特异性无差异；mRT—PCR和胶体金法均可用于人群流感鼻咽拭标本流感病毒快速检测，特别是聚集性暴发流感患者，建议采样时标本数要适当放大2～3倍。mRT—PCR可进一步确定流感病毒的H1和H3亚型。Objective To research the sensitivity and specificity of multiplex reverse transcription-polymerase chain reaction （mRT-PCR） and colloidal gold rapid test for detection of influenza virus. Methods The samples being identified as influenza virus A/H1, A/H3 and B by cell culture and hemagglutinafion inhibition （HI） test were determined by mRT-PCR and colloidal gold rapid test in 78 positive samples and 40 negative samples of nasopharynx swabs. Additionally, the sensitivity was determined in the samples of influenza virus H1 , H3 and B diluted from 101 to 105 TCID50/0. lmL. Results The sensitivity of mRT-PCR to influenza virus A was 44.8% , and its specificity to influenza virus A was 97.5%. The sensitivity of mRT-PCR to influenza virus B was 20.0% , and its specificity to influenza virus B was 100.0%. The sensitivity of colloidal gold rapid test to influenza virus A was 44.8% , and its specificity tn influenza virus A was 100.0%. The sensitivity of colloidal gold rapid test to influenza virus B was 5.0% , and its specificity to influenza virus B was 97.5%. mRT-PCR and colloidal gold rapid test detected 4 samples of A/H1, 4 samples of A/H3 and 3 samples of B being identified by MDCK cell culture, and the positive coincidence and type coincidence of the two tests were 100.0%. The sensitivity of mRT-PCR to influenza virus was 10^3 TCID50/0. 1 mL, and the sensitivity of colloidal gold rapid test to influenza virus was 103 TCID50/0. 1 mL. Conclusions There is no statistical difference in the sensitivity and specificity of mRT-PCR and colloidal gold rapid test. Both mRT-PCR and colloidal gold rapid test can be used in rapid detection of nasopharynx swabs samples of influenza virus, especially in the cluster of patients with explosive influenza virus, but the quantity of sample should be increased 2 to 3 times in the course of sampling. The tests of mRT-PCR can identify subtype H1 and H3 of influenza virus.

关 键 词：流感病毒 分离 多重逆转录聚合酶链反应 胶体金法 

分 类 号：R373.1[医药卫生—病原生物学]