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.

人类非洲锥虫病（HAT）是由锥虫亚种引起的潜在致命寄生虫感染。由采摘苍蝇传播的罗德西森。 HAT监视依赖于主动和被动的病例检测，并且在许多焦点中几乎消除了案例数，需要简单且可持续的筛选方法。在这里，我们描述了一种新型QPCR测定法的原理应用证明，旨在检测采集中的两个人类致病性锥体物种。对96种野生捕获的采采样进行了测定评估，以前被认为是布鲁西t. s阳性的。 l。其中两个对T。b。罗得教。发现该测定法具有高度特异性，与非目标锥虫物种无交叉反应性，检测的测定极限为104杆/mL。 QPCR成功识别了三个T. b。 Rhodesiense阳性果蝇与参考PCR一致。当筛选t. brucei s的致病亚种时，该测定法提供了运行多个PCR的替代方法。 l。并在不到2小时的时间内产生定量结果。这种方法可以提供一种简单有效的方法，用于筛选已知HAT焦点中的大量采采蝇，在有变形或受HAT分布的变化威胁的领域中。