中国人兽共患病学报2012，Vol.28Issue(2)：135-138,4.
梅毒螺旋体TpN47基因分析及实时荧光定量PCR检测方法的建立
Sequences analysis on TpN47 genes from different Treponema pallidum strains and the application of real-time fluorescent quantitative PCR for detection
摘要
Abstract
To establish a real time quantitative PCR to detect T. Pallidum based on outer membrane protein coding gene (TpN47) , the structure domains and transmembrane regions were analyzed through on line database of NCBI, Pfam and TM HMM Server v. 2. 0. The primers were worked out according to the DNA sequence of TpN47. The TpN47 gene was amplified from the total cell DNA of T. Pallidum by PCR, and then inserted into the cloning vector pMD18 T. The sequence of inserted fragment was sequenced, and the T. Pallidum sequences from different T. Pallidum strains were compared through DNAS TAR. Quantitative PCR assay based on TpN47 gene for quick detection of T. Pallidum in serum from people infected by T. Pallidum was conducted. The prediction result suggested that TpN47 located at the outside of outer membrane. The similari ties of nucleotide sequences of TpN47 genes from different T. Pallidum strains were above 99%. The detection result of real time fluorescent quantitative PCR were positive for all the specimens of serum collected from the T. Pallidum infected people. The conserved TpN47 genes were presented in different strains of T. Pallidum. In concluson the real time fluorescent quanti tative PCR has been established for detecting T. Pallidum and its advantages including quickness, stability, sensitivity and specificity, which indicates that this method could be used for clinical laboratory diagnosis of T. Pallidum infection.关键词
梅毒螺旋体/TpN47基因/荧光定量PCRKey words
Treponema pallidum/ TpN47 gene/fluorescent qPCR分类
医药卫生引用本文复制引用
王莹,丁肖青,朱胜,楼宏强..梅毒螺旋体TpN47基因分析及实时荧光定量PCR检测方法的建立[J].中国人兽共患病学报,2012,28(2):135-138,4.基金项目
义乌市科研项目(No.10-3-08)资助 （No.10-3-08）
