梅毒螺旋体荧光定量PCR检测方法的建立

The construction of quantitative PCR method for treponema pallidum

目的建立基于TPN47基因检测血液中梅毒螺旋体的荧光定量PCR检测方法。方法根据梅毒螺旋体TPN47基因的参考序列设计引物和探针,对收集到的200份血液样本进行荧光定量PCR检测,并分析其灵敏性和特异性,同时,对扩增片段进行测序确认。结果荧光定量PCR方法可以扩增到梅毒螺旋体TPN47基因区164 bp长DNA片段,此方法特异性和敏感性都很高;基于TPN47基因的荧光定量PCR方法测定,TPPA阳性血清和TPPA阴性血清的符合率分别为99%(198/200)和98%(196/200)。结论以TPN47基因为扩增的荧光定量PCR方法具有灵敏、快速、便宜等优点,适用于梅毒螺旋体临床实验室诊断。

Objective To establish the quantitative PCR method for detecting Treponema pallidum in blood based on TPN47 gene. Methods According to the reference sequences of treponema pallidum TPN47 gene to design the primers and probes. Then 200 blood samples were collected which measure by quantitative PCR method,and the sensitivity and specificity were analyzed,meanwhile the sequences of amplified fragments were confirmed. Results Quantitative PCR method amplified to the 164 bp DNA fragment of TPN47 gene region with treponema pallidum,it had a high sensitivity and specificity. Determination of TPPA positive serum and negative sera compliance rate was 99%（ 99 /100） and 98%（ 98 /100） respectively,which based on Quantitative PCR method of TPN47 gene.Conclusion Quantitative PCR method based on TPN47 gene detection for treponema pallidum has a high sensitivity and specificity,which could be used for clinical laboratory diagnosis.