基于插入元件IS6110的结核分枝杆菌限制性片段长度多态性基因分型

Insertion element IS6110-based restriction fragment length polymorphism genotyping of Mycobacterium tuberculosis.

作者信息

Warren Robin M, van Helden Paul D, Gey van Pittius Nicolaas C

机构信息

Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Health Sciences, DST/NRF Centre of Excellence in Biomedical Tuberculosis Research, Stellenbosch University, PO Box 19063, Tygerberg, South Africa.

出版信息

Methods Mol Biol. 2009;465:353-70. doi: 10.1007/978-1-59745-207-6_24.

DOI:10.1007/978-1-59745-207-6_24

PMID:20560064

Abstract

DNA fingerprinting techniques are based on genome variation and form the basis of molecular epidemiology studies of tuberculosis. A number of markers are in use for the molecular differentiation of Mycobacterium tuberculosis isolates by DNA fingerprinting. One of these markers is the IS6110 insertion element, which may be present in up to 25 copies per M. tuberculosis genome. Variation in both the number and location of the IS6110 elements makes it a very useful marker of strain genotype. IS6110-based DNA fingerprinting is globally considered as the reference genotyping technique for M. tuberculosis isolates. This method is based on visualization of restriction fragment length polymorphisms using a labeled probe derived from IS6110. In this chapter, the method of IS6110 DNA fingerprinting is explained in such a way that it can be easily duplicated by molecular epidemiologists and will give reproducible results.

摘要

DNA指纹技术基于基因组变异，构成了结核病分子流行病学研究的基础。目前有多种标记物用于通过DNA指纹技术对结核分枝杆菌分离株进行分子鉴别。其中一种标记物是IS6110插入元件，每个结核分枝杆菌基因组中可能存在多达25个拷贝。IS6110元件数量和位置的变异使其成为菌株基因型的一个非常有用的标记物。基于IS6110的DNA指纹技术在全球范围内被视为结核分枝杆菌分离株的参考基因分型技术。该方法基于使用源自IS6110的标记探针来可视化限制性片段长度多态性。在本章中，IS6110 DNA指纹技术的方法将以一种易于分子流行病学家重复且能给出可重复结果的方式进行解释。