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。
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.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2004年第10期603-605,共3页
Chinese Journal of Dermatology
