梅毒感染诊断中Tp0259-PCR方法的建立  

Establishment of Tp0259-PCR for diagnosing infection of treponema pallidum

作　　者：杨长顺[1] 王三虎[1] 周秀萍 

机构地区：[1]怀化市第一人民医院检验科,湖南怀化418000 [2]湖南医药学院,湖南怀化418000

出　　处：《国际检验医学杂志》2015年第15期2232-2233,共2页International Journal of Laboratory Medicine

基　　金：怀化市科技局课题(怀财企指[2013]4号1-13);湖南省教育厅课题(11C1014)

摘　　要：目的建立聚合酶链反应(PCR)方法扩增梅毒螺旋体Tp0259基因,为进一步探讨其在梅毒感染诊断中的作用提供依据。方法从Genbank获取Tp0259基因序列,生物信息学分析,设计特异性引物。收集Tp Nichols株感染组和对照组的新西兰兔睾丸标本,提取全基因组DNA,通过优化PCR扩增条件寻找最佳反应体系和反应条件,PCR扩增Tp0259的编码基因,同时通过新构建扩增方法检测泌尿生殖道感染常见病原微生物。结果通过PCR扩增及琼脂糖凝胶电泳,只有感染梅毒螺旋体的新西兰兔标本才能扩增出特异性条带,条带位置约为646bp,其他病原微生物未见扩增条带。结论检测Tp0259基因能用于诊断梅毒螺旋体的感染。Objective To establish a method for detection Tp0259 gene by PCR .To provide basis of diagnosis infection of trepo‐nema pallidum by detection Tp0259 gene .Methods Gene sequences of Tp0259 was obtained from Genebank and analysis by bioin‐formatics .Specific primers were designed .The whole genome was extracted from testis specimens of the New Zealand rabbits infec‐ted by treponema pallidum and control groups .The gene of Tp0259 was amplified by PCR under the optimization reaction condi‐tions which was selected by many tests ,using the new construction of amplification method to detect the common pathogenic micro‐organism in urinary and reproductive tract infection .Results Specific bands of 646 bp was appeared only in New Zealand rabbits which was infected by treponema pallidum ,while none was in the control group and other pathogenic microorganisms .Conclusion Detection the Tp0259 gene could be used to diagnose infection of treponema pallidum .

关 键 词：梅毒螺旋体 聚合酶链反应 基因 

分 类 号：R759.1[医药卫生—皮肤病学与性病学]