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用于鉴定阴道毛滴虫DNA的分子探针。

Molecular probe for identification of Trichomonas vaginalis DNA.

作者信息

Rubino S, Muresu R, Rappelli P, Fiori P L, Rizzu P, Erre G, Cappuccinelli P

机构信息

Istituto di Microbiologia e Virologia, Università degli Studi di Sassari, Italy.

出版信息

J Clin Microbiol. 1991 Apr;29(4):702-6. doi: 10.1128/jcm.29.4.702-706.1991.

Abstract

Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic observation of vaginal discharge, cell culture, and immunological techniques. A 2.3-kb Trichomonas vaginalis DNA fragment present in strains from diverse geographic areas was cloned and used as a probe to detect T. vaginalis DNA in vaginal discharge by a dot blot hybridization technique. This probe was specific for T. vaginalis DNA. It recognized strains from two regions in Italy (Sardinia, Piemonte) and from Mozambique (Africa). In addition, our probe did not cross-react with bacterial (Escherichia coli, Enterococcus spp., group B streptococci, Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, and Lactobacillus spp.), viral (herpes simplex virus type 2), fungal (Candida albicans), protozoan (Entamoeba histolytica, Giardia lamblia, Plasmodium falciparum, Leishmania major, and Leishmania infantum), or human nucleic acids. The probe reacted with Pentatrichomonas hominis and Trichomonas foetus. The limit signal recognized by our probe corresponded to the DNA of 200 T. vaginalis isolates. The 2.3-kb probe was used in a clinical analysis of 98 samples. Of these, 20 samples were found to be positive both with the probe and by cell culture, and only 14 of these were positive by a standard wet mount method.

摘要

滴虫病是世界上传播最广泛的性传播疾病之一。可通过多种方法进行诊断,如直接显微镜观察阴道分泌物、细胞培养和免疫技术。克隆了来自不同地理区域菌株中存在的一个2.3kb阴道毛滴虫DNA片段,并将其用作探针,通过斑点印迹杂交技术检测阴道分泌物中的阴道毛滴虫DNA。该探针对阴道毛滴虫DNA具有特异性。它识别来自意大利两个地区(撒丁岛、皮埃蒙特)和莫桑比克(非洲)的菌株。此外,我们的探针与细菌(大肠杆菌、肠球菌属、B组链球菌、阴道加德纳菌、淋病奈瑟菌、沙眼衣原体和乳杆菌属)、病毒(2型单纯疱疹病毒)、真菌(白色念珠菌)、原生动物(溶组织内阿米巴、蓝氏贾第鞭毛虫、恶性疟原虫、硕大利什曼原虫和婴儿利什曼原虫)或人类核酸均无交叉反应。该探针与口腔毛滴虫和胎儿毛滴虫有反应。我们的探针识别的极限信号相当于200株阴道毛滴虫的DNA。该2.3kb探针用于对98份样本的临床分析。其中,20份样本经探针检测和细胞培养均呈阳性,而其中只有14份通过标准湿片法呈阳性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5928/269856/061f754fa779/jcm00040-0053-a.jpg

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