比较实时荧光定量PCR法和直接荧光免疫法检测人偏肺病毒的价值  

A comparison between real-time quantitative fluorescence polymerase chain reaction assay and direct immunofluorescence assay for diagnosis of human metapneumovirus

作　　者：苗正友[1] 徐营[2] 李平[2] 

机构地区：[1]嘉兴市妇幼保健院产前诊断中心实验室,314001 [2]嘉兴学院医学院组胚教研室

出　　处：《中华检验医学杂志》2011年第1期50-54,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的 比较Q-RT-PCR法与直接荧光免疫法对诊断hMPV的价值.方法 收集嘉兴市妇幼保健院2008年11月至2009年5月因急性呼吸道感染入院治疗的患儿共1 283例,分离hMPV阳性病毒株,进行测序并与荷兰分离株NLD00-1进行序列比对.根据本地区流行的hMPV病毒株序列设计hMPV特异的引物和荧光标记探针,建立应用TaqMan探针的Q-RT-PCR方法.用负压吸引的方法采集鼻咽分泌物标本,标本分别用Q-RT-PCR和FITC标记的病毒特异性单克隆抗体直接荧光免疫法进行检测,并对两种方法的检测结果进行McNemar test一致性检验.结果 1283份标本同时进行Q-RT-PCR和直接荧光免疫法两种方法检测,Q-RT-PCR法检出59例阳性,直接荧光免疫法检出55例阳性,两者同时为阳性者52例,Q-RT-PCR法阳性而直接荧光免疫法阴性7例,Q-RT-PCR阴性而直接荧光免疫法阳性3例.如果以Q-RT-PCR法结果为金标准,直接荧光免疫法检测敏感度为88.1%,特异度为99.8%,阳性预测值为94.5%,阴性预测值为99.4%,准确性为99.2%.McNemar test一致性检验,两种方法检测阳性率差异无统计学意义（χ2=0.9,P＞0.05）.结论直接荧光免疫法对hMPV临床诊断价值接近于Q-RT-PCR.Objectives To evaluate the diagnostic value of real-time quantitative fluorescence polymerase chain reaction（ Q-RT-PCR ） assay and immunofluorescence assay for diagnosis of hMPV. Methods Totally 1 283 children with acute respiratory infection admitted in Jiaxing Maternity and Child Health Care Hospital for treatment from November 2008 to May 2009 were recruited in this study. The hMPV positive stains were separated and sequenced in this area. The sequences between the local hMPV stains and Holland stains NLD00-1 were compared. The specific primers and fluorescent probe were designed according to the sequence of epidemic hMPV strain. The Taqman methodology was applied in Q-RT-PCR. Negative pressure suction was used to acquire nasopharyngeal secretions specimens. Both Q-RT-PCR and immunofluorescence with FITC labeled monoclonal antibody were used to analyze them, respectively. The McNemar, test was applied to analyze the correlation between the two methods. Results Totally 1 283 specimens were analyzed with Q-RT-PCR and immunofiuorescence simultaneously. Q-RT-PCR analysis showed there were 59 cases positive. Immunofluorescence analysis showed there were 55 cases positive. Fifty-two cases were positive in both assays. There were 7 cases positive in Q-RT-PCR assay but negative in immunofluorescence assay and 3 cases negative in Q-RT-PCR assay but positive in immunofluorescence assay. If Q-RT-PCR method was set as the golden standard, the sensitivity and specificity for immunofluorescence detection method were 88. 1%and 99. 8%, respectively. Positive predictive value and negative predictive value were 94. 5% and 99. 4%,respectively. There was no significant difference （ χ2= 0. 9, P ＞ 0. 05 ） by McNemar＇ test between the two methods. Conclusion The diagnostic value of immunofluorescence assay is close to Q-RT-PCR assay.

关 键 词：变性肺病毒 荧光抗体技术 直接 逆转录聚合酶链反应 

分 类 号：R72[医药卫生—儿科] R4[医药卫生—临床医学]