自身淬灭荧光技术定量检测临床分离的淋病奈瑟菌DNA

Detection of Neisseria gonorrhoeae DNA by fluorescent quantitative PCR using self-quenched primer technology

目的 探讨自身淬灭荧光定量聚合酶链反应（FQ-PCR）检测临床分离的淋病奈瑟菌DNA的临床应用价值。方法 以建立的淋病奈瑟菌自身淬灭FQ-PCR方法为基础，对59例疑为淋病奈瑟菌感染患者的标本进行检测，并与分离培养法进行分析比较。结果自身淬灭FQ-PCR标准品浓度为10^2～10^7拷贝／μ时，方法能够保持良好的线性关系，标准曲线为：Y=-0．273X＋10．781，相关系数为0．9895。自身淬灭FQ-PCR阳性检出率为64．41％，高于培养法52．54％（χ^2=5．14，P〈0．05）；两者特异性均为100％。结论 自身淬灭FQ-PCR用于临床分离的淋病奈瑟菌DNA检测，敏感性、特异性均高，对淋病的诊断有较高价值，实现了PCR扩增、荧光探针杂交及检测一体化，简便省时。

Objective To evaluate the effects of fluorescent quantitative PCR （FQ-PCR） using self-quenched primer technology in detecting the Neisseria gonorrhoeae DNA of clinical samples. Methods FQ-PCR basing on self-quenched primer technology was applied to detect 59 clinical specimens from patients with suspected Neisseria gonorrhoeae infection, and the result was compared with bacterial culturing. Results When standard concentration of FQ-PCR using self-quenched primer was 10^2- 10^7 copes/μL, the method could maintain good linear relationship, the standard curve was：Y= - 0. 273X ＋ 10. 781, the correlation coefficient was 0. 9895. The positive rate of FQ- PCR and bacterial culture was 64. 41 % and 52. 54% respectively（χ^2 = 5.14, P〈0.05）, specificity of both methods were 100%. Conclusion FQ-PCR basing on self-quenched primer assay is sensitive and specific, which is valuable for gene diagnosis of Neisseria gonorrhoeae infection.