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用于廉价检测结核分枝杆菌单核苷酸多态性的反向线探针分析

Reverse line probe assay for cheap detection of Single Nucleotide Polymorphisms in Mycobacterium tuberculosis.

作者信息

Yasmin Memona, Refregier Guislaine, Siddiqui Rubina Tabassum, Iqbal Rizwan, Abbasi Shahid Ahmad, Tahseen Sabira

机构信息

Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O.Box No. 577, Jhang Road, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.

Institut de Génétique et Microbiologie, UMR8621, CNRS - Univ. Paris-Sud, Universud, Campus d'Orsay, F-91405 Orsay-Cedex, France.

出版信息

Tuberculosis (Edinb). 2018 May;110:52-55. doi: 10.1016/j.tube.2018.03.007. Epub 2018 Mar 26.

Abstract

More and more Single Nucleotide Polymosrphisms of interest among pathogenic organisms are described with the advent of Whole Genome Sequencing but WGS approach is still too expensive, time consuming, and relying on bioinformatical means that are not available in many developing countries. This study presents a low-cost reverse hybridization line probe technique for detecting SNPs in Mycobacterium tuberculosis. The proposed test is able to detect mutations in the RRDR of rpoB gene in M. tuberculosis with specificity and sensitivity of 98% and 100%, respectively and for an average cost of less than €3 per sample. The technique proved efficient not only on pure DNA samples extracted from culture isolates but also on crude extracts from clinical samples. The flexibility of the platform allows to get it transformed to any kind of test detection, hence, building a bridge between rich countries performing SNP discovery and countries with high burden that can target these SNPs on the collected samples.

摘要

随着全基因组测序技术的出现,越来越多致病生物中感兴趣的单核苷酸多态性被描述出来,但全基因组测序方法仍然过于昂贵、耗时,并且依赖于许多发展中国家无法获得的生物信息学手段。本研究提出了一种用于检测结核分枝杆菌单核苷酸多态性的低成本反向杂交线探针技术。所提出的检测方法能够检测结核分枝杆菌rpoB基因RRDR区域的突变,特异性和敏感性分别为98%和100%,每个样本的平均成本低于3欧元。该技术不仅在从培养分离物中提取的纯DNA样本上证明是有效的,而且在临床样本的粗提物上也有效。该平台的灵活性允许将其转化为任何类型的检测,因此,在进行单核苷酸多态性发现的富国与能够对收集样本中的这些单核苷酸多态性进行靶向检测的高负担国家之间架起了一座桥梁。

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