CAJ | 학술논문

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5×102～5×108 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-18例（占18.2%），标本病毒载量为8.5546×106 copies/ml，HSV-236例（占81.8%），病毒载量为1.9861×106 copies/ml。FQ-PCR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。
목적：평개형광정량취합매련반응（FQ-PCR）재생식기포진（GH）진단화분형중적응용。방법이단순포진병독(HSV)-1 DNA다취매기인화HSV-2당단백D기인위파기인구，설계합성정향、반향인물화탐침，분별대HSV-1화HSV-2진행FQ-PCR검측，우화반응체계，진행방법학평개，병대의사GH환자적생식기면식자표본채용FQ-PCR진행검측화분형。결과건립적FQ-PCR대HSV검측화분형구유특이성；선성범위호（표준품적농도위5×102～5×108 copies/ml，r=0.998）；령민도체도5×102 copies/ml；중복성교호，비내CV치위2.29%，비간CV치위4.76%。186례환자HSV양성44례，양성솔위23.7%（44/186）；기중44례양성자중HSV-18례（점18.2%），표본병독재량위8.5546×106 copies/ml，HSV-236례（점81.8%），병독재량위1.9861×106 copies/ml。FQ-PCR여PCR법적HSV검출솔여분형검측결과일치。결론건립적FQ-PCR방법구유고특이성화민감성，분형、정량준학，방법쾌속、간편，가용우GH적진단화분형。
Objective To estimate the value of the lfuorescence 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 speciifcity for detection and typing of HSV, with good linear range (5×102-5×108 copies/ml, r=0.998), and a sensitivity of 5×102 copies/ml, as well as good reproducibility (infra-assay coefifcients of variation was 2.29%and inter-assay coefifcients 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 8.5546×106 copies/ml and 36 (81.2%) were positive for HSV-2 with a viral load of 1.9861×106 copies/ml. Conclusion FQ-PCR is a speciifc, sensitive and rapid method for the detection and typing of HSV, which can be used in clinical diagnosis and typing of genital herpes.