A pilot study demonstrating the identification of Trypanosoma brucei gambiense and T. b. rhodesiense in vectors using a multiplexed high-resolution me

一项初步研究证明布氏冈比亚锥虫和 T. b. 的鉴定。

基本信息

批准号：
1964757
负责人：
金额：
--
依托单位：
Liverpool School of Tropical Medicine
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2017
资助国家：
英国
起止时间：
2017 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-1964757
关键词：
pilot study demonstrating identification Trypanosoma

项目摘要

Human African Trypanosomiasis (HAT) is a potentially fatal parasitic infection caused by the trypanosome sub-species Trypanosoma brucei gambiense and T. b. rhodesiense transmitted by tsetse flies. HAT surveillance is reliant on active and passive case detection and with case numbers nearing elimination in many foci there is a need for simple and sustainable screening methods. Here, we describe a proof of principle application of a novel qPCR assay designed to detect both human pathogenic trypanosome species in tsetse. Assay evaluation was carried out on 96 wild-caught tsetse previously identified to be positive for T. brucei s. l. of which two were known to be positive for T. b. rhodesiense. The assay was found to be highly specific with no cross-reactivity with non-target trypanosome species and the assay limit of detection was 104 tryps/mL. The qPCR successfully identified three T. b. rhodesiense positive flies, in agreement with the reference PCRs. This assay provides an alternative to running multiple PCRs when screening for pathogenic sub-species of T. brucei s. l. and produces quantitative results in less than 2 hours. This method could provide a component of a simple and efficient method of screening large numbers of tsetse flies in known HAT foci, in areas at risk of recrudescence or threatened by the changing distribution of HAT.

非洲人类锥虫病 (HAT) 是一种潜在致命的寄生虫感染，由锥虫亚种布氏冈比亚锥虫 (Trypanosoma brucei gambiense) 和 T. b. 引起。罗德西亚病由采采蝇传播。 HAT 监测依赖于主动和被动病例检测，随着许多重点地区的病例数量接近消除，需要简单且可持续的筛查方法。在这里，我们描述了一种新型 qPCR 测定的原理应用证明，该测定旨在检测采采蝇中的两种人类致病性锥虫物种。对先前鉴定为布氏锥虫呈阳性的 96 只野生捕获采采蝇进行了测定评估。湖其中两个已知 T. b. 呈阳性。罗德西亚。该检测具有高度特异性，与非目标锥虫物种没有交叉反应性，检测限为 104 锥虫/mL。 qPCR 成功鉴定了三个 T。罗德西亚果蝇呈阳性，与参考 PCR 一致。在筛选 T. brucei 致病亚种时，该测定法提供了运行多重 PCR 的替代方法。湖并在不到 2 小时内产生定量结果。该方法可以提供一种简单而有效的方法的组成部分，用于在已知的 HAT 病灶、有复发风险或受到 HAT 分布变化威胁的区域中筛查大量采采蝇。