Genetic subtyping of human immunodeficiency virus using a heteroduplex mobility assay.
Genetic subtyping of human immunodeficiency virus using a heteroduplex mobility assay.
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DOI：
10.1101/gr.4.5.s202
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发表时间：
1995-04
期刊：
影响因子：
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通讯作者：
Eric Delwart;B. Herring;Allen G. Rodrigo;James I. Mullins
中科院分区：
文献类型：
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作者：
Eric Delwart;B. Herring;Allen G. Rodrigo;James I. Mullins
同一种微生物的不同菌株往往表现出不同的特性。在越来越多的情况下，表型特征与遗传标记的联系使得复杂的生物测定可以被基因分型所取代。这种遗传变异的检测已经被PCR彻底改变，它允许即使是最复杂的基因组的片段在几个小时内以基本上纯的形式分离出来。然后可以通过PCR产物的直接测序来确定基因片段之间的差异。为了进一步加速，并由此将遗传筛选测定扩展到更多数量的样品，已经开发了多种非测序方法，其比DNA测序更简单，通常需要不太复杂的设备，并且成本更低。例如，异源双链体分析已用于医学遗传学领域（1-7）和用于检测人类群体中的遗传多态性。（8 '9）这些方法涉及使用非变性以及轻微变性的凝胶电泳条件。乙烯基聚合物凝胶也已用于诊断原癌基因内的突变。（12-14~非测序遗传分析方法也可以促进复杂和快速进化的遗传系统，如RNA病毒的研究。最近描述的异源双链迁移率测定，~现在用于分类的HIV-1病毒株的遗传亚型，在这里介绍。这些测定法也应该容易适用于其他高度可变的微生物的分析。
Different strains of the same" species" of microorganism often display distinctive properties. In a growing number of cases, linkage of phenotypic traits with genetic markers is allowing complicated biological assays to be replaced by genetic typing. The detection of such genetic variation has been revolutionized by the PCR, which allows fragments of even the most complex genomes to be isolated in an essentially pure form in a matter of hours. Differences between gene segments can then be determined by direct sequencing of the PCR product. To further expedite, and thereby to extend genetic screening assays to greater numbers of samples, multiple nonsequencing methods have been developed that are simpler, typically require less complex apparatus, and are of lower cost than DNA sequencing. For example, heteroduplex analysis has been used in the field of medical genetics (1-7) and for the detection of genetic polymorphisms in human populations.(8'9) These methods have involved use of nondenaturing as well as slightly denaturing gel electrophoresis conditions.(1~ Vinyl polymer gels have also been used for the diagnosis of mutations within proto-oncogenes.(12-14~ Nonsequencing genetic analysis methods can also facilitate study of complex and rapidly evolving genetic systems such as RNA viruses. Recently described heteroduplex mobility assays,~ now used to classify HIV-1 strains into genetic subtypes, are presented here. These assays should also be readily applicable to the analysis of other highly variable microorganisms.