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筛选针对阴道毛滴虫粘附蛋白的适体用于诊断应用

Aptamer Selection against a Trichomonas vaginalis Adhesion Protein for Diagnostic Applications.

作者信息

Espiritu Christian Adam L, Justo Christine Aubrey C, Rubio Miriam Jauset, Svobodova Marketa, Bashammakh Abdulaziz S, Alyoubi Abdulrahman O, Rivera Windell L, Rollon Analiza P, O'Sullivan Ciara K

机构信息

Interfibio Group, Departament d'Enginyeria Química , Universitat Rovira i Virgili , Avinguda Països Catalans, 26 , Tarragona 43007 , Spain.

Department of Chemistry, Faculty of Science , King Abdulaziz University , Jeddah 21589 , Kingdom of Saudi Arabia.

出版信息

ACS Infect Dis. 2018 Sep 14;4(9):1306-1315. doi: 10.1021/acsinfecdis.8b00065. Epub 2018 Jul 20.

Abstract

Trichomoniasis, caused by Trichomonas vaginalis, is the leading nonviral sexually transmitted infection worldwide. We report the selection of a DNA aptamer against a T. vaginalis adhesion protein, AP65, using a microtiter plate-based in vitro combinatorial chemistry process termed systematic evolution of ligands by exponential enrichment. The enriched library pool was sequenced by next-generation sequencing, and several aptamer candidates with high affinity and specificity were identified. The aptamer with the highest affinity and specificity had a K in the low nanomolar range, as confirmed by three different techniques: surface plasmon resonance, enzyme-linked aptamer assay, and biolayer interferometry. The selected aptamer was demonstrated to have a high specificity to the AP65 protein and to T. vaginalis cells with no cross-reactivity to other enteric and urogenital microorganisms. Current work is focused on the development of inexpensive and easy-to-use aptamer-based diagnostic assays for the reliable and rapid detection of T. vaginalis in vaginal swabs.

摘要

由阴道毛滴虫引起的滴虫病是全球主要的非病毒性性传播感染。我们报告了通过一种基于微孔板的体外组合化学过程(称为指数富集配体系统进化)来筛选针对阴道毛滴虫粘附蛋白AP65的DNA适配体。通过下一代测序对富集的文库池进行测序,并鉴定出几种具有高亲和力和特异性的适配体候选物。通过表面等离子体共振、酶联适配体测定和生物层干涉术这三种不同技术证实,具有最高亲和力和特异性的适配体的K值处于低纳摩尔范围。所选适配体对AP65蛋白和阴道毛滴虫细胞具有高特异性,对其他肠道和泌尿生殖微生物无交叉反应。目前的工作重点是开发基于适配体的廉价且易于使用的诊断检测方法,用于可靠、快速地检测阴道拭子中的阴道毛滴虫。

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