中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (10): 603-605.

• 论著 • 上一篇    下一篇

PCR在阴道毛滴虫检测中的引物筛选和条件优化

徐敏, 尹跃平, 余艳华, 刘爱英, 王书崎   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042
  • 收稿日期:2003-10-30 出版日期:2004-10-15 发布日期:2004-10-15
  • 通讯作者: 尹跃平,E-mail:ypyinc@jlonlion.com E-mail:ypyinc@jlonlion.com

Optimization of PCR Components and Selection of Its Primers for Detection of Trichomonas Vaginalis

XU Min, YIN Yue-ping, YU Yan-hua, LIU Ai-ying, WANG Shu-qi   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2003-10-30 Online:2004-10-15 Published:2004-10-15

摘要: 目的 探讨PCR检测阴道毛滴虫的引物筛选和条件优化。方法 从文献中筛选TVA5-TVA6、TV1-TV2、TVK3-TVK73对引物进行比较,每种PCR对引物、Mg2+、dNTPs、Taq酶4个因素,每个因素取3个浓度水平,使用Taguchi法进行优化,筛选出每种PCR的较佳反应条件。在较佳反应条件下,比较3种PCR的敏感性。用培养法及PCR法检测阴道拭子标本,初步评价敏感性最高的PCR方法。结果 3种PCR法均具有较高的特异性,TVK3-TVK7PCR的敏感性最高。25份临床拭子标本中,培养法检测出7份阳性标本,TVK3-TVK7PCR方法检测出8份,所有培养法检测阳性的标本经PCR方法检测的结果均为阳性。结论 TVK3-TVK7PCR具有高度敏感性和特异性,是一种检测阴道毛滴虫感染的有效方法。

关键词: 毛滴虫, 阴道, 聚合酶链反应

Abstract: Objective To screen primers used in polymerase chain reaction (PCR) for detecting Trichomonas vaginalis. Methods Three pairs of PCR primer reported in the literatures (TVA5-TVA6, TV1-TV2 and TVK3-TVK7) were screened. For each PCR, four components, including primers, Mg2+, dNTPs and Taq polymerase, were optimized using Taguchi methods to determine the optimal PCR conditions. With the optimal conditions, the sensitivities of three PCR were compared. Vaginal swabs were collected to detect Trichomonas vaginalis by culture and PCR, and the PCR with highest sensitivity was evaluated. Results All three PCR were of high specificity, and the PCR with primers of TVK3-TVK7 had the highest sensitivity. Of 25 clinical vaginal swabs, T. vaginalis was detected in 7 samples by the culture, however, it was detected in 8 samples by the PCR. All culture-positive samples were also positive by PCR. Conclusions The PCR with the primers of TVK3-TVK7 is highly sensitive and specific, which could be useful to detect T. vaginalis in vaginal swab samples.

Key words: Trichomonas vaginalis, Polymerase chain reaction