半巢式聚合酶链反应快速诊断单纯疱疹病毒性脑炎及病毒分型

Rapid Diagnosis of Herpes Simplex Encephalitis and Viral Genotyping by Semi-Nested Polymerase Chain Reaction

目的:进一步提高PCR诊断方法的特异性和敏感性。方法:在单纯疱疹病毒(HSV)TK基因区设计了一对外层公用引物和两个内层分型引物,建立了半巢式PCR检测和分型HSV的方法。用该方法检测了天津地区64例病毒性脑炎患者(病脑组),和46例其他中枢神经系统疾病患者(对照组)的脑脊液(CSF)标本。结果:病脑组阳性率为15.63%(10/64),其中9例为HSV-1感染,1例为HSV-2感染。对照组阳性率为2.17%(1/46)。结论:本方法在发病后第1天即可测出CSF中的HSV DNA,敏感性为0.01TCID_(50)。

Purpose and Methods: A semi-nested polymerase chain reaction (PCR), flanking a region of the HSV thymidine kinase (TK)gene where a pair of outer public primers and two typing primers were selected, was set up to detect and differentiate HSV-1 and HSV-2. Using the method, 64 CSF samples from patients with viral encephalitis were examined, and 46 CSF samples from patients with other central nervous system diseases were examined as control. Results:The positive rate of viral encephalitis group was 15.63%(10/64),including nine patients suffered from HSV-1 and one patient infected by HSV-2; whereas the positive rate of control group was 2.17% (1 /46). Conclusion: The HSV DNA in patient' s CSF could be detected clearly by the semi-nested PCR assay established in TK gene region as early as one day after onset, and the sensitivity reached O.OlTCID50. Therefore,it may be of higher available value for clinical diagnosis of herpes simplex encephalitis.