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A 基因组串联重复卫星 DNA 序列作为一种分子标记,用于诊断恰加斯病的 LAMP 检测。

A Genome Tandem Repetitive Satellite DNA Sequence as a Molecular Marker for a LAMP Assay for Diagnosing Chagas' Disease.

机构信息

Animal Science Faculty, Universidad de Ciencias Aplicadas y Ambientales (U.D.C.A), 111166 Bogotá, Colombia.

Infectious and Tropical Diseases Research Group (e-INTRO), Biomedical Research Institute of Salamanca, Research Centre for Tropical Diseases at the University of Salamanca (IBSAL-CIETUS), Faculty of Pharmacy, University of Salamanca, 37007 Salamanca, Spain.

出版信息

Dis Markers. 2020 Feb 22;2020:8074314. doi: 10.1155/2020/8074314. eCollection 2020.

Abstract

Chagas' disease is a neglected tropical disease caused by which is endemic throughout Latin America and is spread by worldwide migration. Diagnosis is currently limited to serological and molecular techniques having variations regarding their sensitivity and specificity. This work was aimed at developing a new sensitive, applicable, and cost-effective molecular diagnosis technique for loop-mediated isothermal amplification-based detection of (Tc-LAMP). The results led to determining a highly homologous satellite repeat region (231 bp) among parasite strains as a molecular marker for diagnosing the disease. Tc-LAMP was performed correctly for detecting parasite DNA (5 fg for the CL Brener strain and 50 fg for the DM28, TcVI, and TcI strains). Assay results proved negative for DNA from 16 helminth species and 7 protozoa, including spp. Tc-LAMP based on the highly repeated satellite region is thus proposed as an important alternative for diagnosing infection, overcoming other methods' limitations such as their analytic capability, speed, and requiring specialized equipment or highly trained personnel. Tc-LAMP could be easily adapted for point-of-care testing in areas having limited resources.

摘要

恰加斯病是一种被忽视的热带病,由 引起,该疾病在拉丁美洲各地流行,并通过全球移民传播。目前的诊断仅限于血清学和分子技术,这些技术在敏感性和特异性方面存在差异。本工作旨在开发一种新的敏感、适用且具有成本效益的分子诊断技术,用于基于环介导等温扩增的检测 (Tc-LAMP)。研究结果确定了寄生虫株之间高度同源的卫星重复区(231bp)作为诊断该病的分子标记。Tc-LAMP 可正确检测寄生虫 DNA(CL Brener 株为 5fg,DM28、TcVI 和 TcI 株为 50fg)。该检测方法对来自 16 种蠕虫和 7 种原生动物的 DNA 检测结果均为阴性,包括 spp. 因此,基于高度重复的 卫星区的 Tc-LAMP 被提议作为诊断 感染的重要替代方法,克服了其他方法在分析能力、速度以及需要专用设备或经过高度训练的人员等方面的局限性。Tc-LAMP 可以很容易地适应资源有限地区的即时检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/7060435/08796e657c67/DM2020-8074314.001.jpg

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