巢式PCR检测梅毒螺旋体DNA方法.pdfVIP

·188 · CHINA TROPICAL MEDICINE Vol . 12 No .2 February 2012 中国热带医学2012 年第12 卷第2 期 ·论 著· 巢式PCR 检测梅毒螺旋体DNA 方法研究 王佃鹏，易娟，张艳芳，汤海燕，邓丽丹，罗斌，李培茂，张志敏，童向东 摘要：目的 建立一种血清梅毒螺旋体DNA 提取及PCR 鉴定方法。 方法 基于化学热裂解法和柱式核酸提取 原理进行血清梅毒螺旋体DNA 提取。根据文献筛选优化引物建立巢式PCR 检测体系，并对实验方法的特异性、抗干扰 性、检测梯度进行评价。 结果 建立的实验方法特异性好、抗干扰性强，对TRUST 为 1：8 以上的标本检测全部为阳 性。 结论 提取梅毒螺旋体DNA 及PCR 鉴定有助于梅毒的快速诊断，对于血液中梅毒螺旋体增殖能力判断具有重 大潜在应用价值。 关键词：梅毒螺旋体；DNA 提取；巢式PCR 中图分类号：R377.1 文献标识码：A 文章编号：1009- 9727（2012）2- 188- 03 Methodological study on detection of Treponema pallidum DNA with nested PCR. WANG Dian- peng，YI Juan ， ZHANG Yan- fang，et al. （Shenzhen Hospital for Occupational Disease Prevention and Treatment; Shenzhen 518001，Guangdong， P. R. China） Abstract ：Objective To establish a method for detecting serum Treponema pallidum DNA with nested PCR detection. Methods Serum Treponema pallidum DNA was extracted on the basis of chemical pyrolysis method and principles of nucleic acid extraction column. According to the literature，primers were screened and optimized and established the nested PCR detection system. The fingdings of specificity，anti- interference and gradient test were evaluated，and the clinical specimens were tested. Results This experimental method was extremely specific and showed a strong anti- interference features. The clinical detection showed that the samples with syphilis toluidine red untreated serum test （TRUST）titre 1:8 or above were all positive. Conclusion Both serum Treponema pallidum DNA extraction and nested PCR detection allow rapidly diagnosis of syphilis and that show great potential for confirming proliferative ability of Treponema pallidum in blood. Key words: Treponema p