荧光定量聚合酶链反应在生殖器疱疹诊断和分型中的应用  

Application of the fluorescence quantitative polymerase chain reaction in the diagnosis and typing of genital herpes

作　　者：刘军连[1] 杜海平[1] 司少艳[2] 徐冰心[2] 敬华[3] 吕世超[1] 易勇[3] 

机构地区：[1]解放军第306医院皮肤科,北京100101 [2]解放军第306医院特种医学实验研究中心 [3]解放军第306医院检验科

出　　处：《实用皮肤病学杂志》2014年第4期250-252,共3页Journal of Practical Dermatology

基　　金：国家自然科学基金资助项目(30872284;81071402)

摘　　要：目的评价荧光定量聚合酶链反应（FQ-PCR）在生殖器疱疹（GH）诊断和分型中的应用。方法以单纯疱疹病毒（HSV）-1 DNA多聚酶基因和HSV-2糖蛋白D基因为靶基因区,设计合成正向、反向引物和探针,分别对HSV-1和HSV-2进行FQ-PCR检测,优化反应体系,进行方法学评价,并对疑似GH患者的生殖器棉拭子标本采用FQ-PCR进行检测和分型。结果建立的FQ-PCR对HSV检测和分型具有特异性;线性范围好（标准品的浓度为5×10^2～5×10^8 copies/ml,r=0.998）;灵敏度达到5×102 copies/ml;重复性较好,批内CV值为2.29%,批间CV值为4.76%。186例患者HSV阳性44例,阳性率为23.7%（44/186）;其中44例阳性者中HSV-1 8例（占18.2%）,标本病毒载量为8.5546×10^6 copies/ml,HSV-2 36例（占81.8%）,病毒载量为1.9 861×10^6 copies/ml。FQPCR与PCR法的HSV检出率与分型检测结果一致。结论建立的FQ-PCR方法具有高特异性和敏感性,分型、定量准确,方法快速、简便,可用于GH的诊断和分型。Objective To estimate the value of the fl uorescence quantitative polymerase chain reaction（FQ-PCR） method in the diagnosis and typing of genital herpes. Methods By employing the FQ-PCR technique, the primers and probes targeted at HSV-1 DNA polymerase gene and HSV-2 glycoprotein D gene fraction were designed and applied to amplify DNA from HSV-1 or HSV-2. Then the PCR reaction system was optimized and evaluated. Swab specimens from suspected patients with genital herpes were detected for HSV by FQ-PCR. Results The FQ-PCR assay showed good specifi city for detection and typing of HSV, with good linear range（5×10^2～5×10^8 copies/ml, r=0.998）, and a sensitivity of 5×10^2 copies/ml, as well as good reproducibility（infra-assay coeffi cients of variation was 2.29% and interassay coeffi cients of variation was 4.76%）. Total 186 swab specimens were tested for HSV by FQ-PCR, the positive rate was 23.7%（44/186）, among the 44 positive specimens, 8（18.2%） were positive for HSV-1 with a viral load of 8.5546×10^6 copies/ml and 36（81.2%） were positive for HSV-2 with a viral load of 1.9861×10^6 copies/ml. Conclusion FQ-PCR is a specifi c, sensitive and rapid method for the detection and typing of HSV, which can be used in clinical diagnosis and typing of genital herpes.

关 键 词：生殖器疱疹 单纯疱疹病毒 荧光定量聚合酶链反应 诊断 分型 

分 类 号：R752.1[医药卫生—皮肤病学与性病学]