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首页> 外文期刊>The Southeast Asian journal of tropical medicine and public health >DEVELOPMENT OF SINGLE-TUBE MULTIPLEX PCR FOR CLASSIFICATION OF MYCOBACTERIUM TUBERCULOSIS LINEAGES BASED ON LARGE SEQUENCE POLYMORPHISMS
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DEVELOPMENT OF SINGLE-TUBE MULTIPLEX PCR FOR CLASSIFICATION OF MYCOBACTERIUM TUBERCULOSIS LINEAGES BASED ON LARGE SEQUENCE POLYMORPHISMS

机译：基于大序列多态性的单管多重PCR用于结核分枝杆菌谱系分类的开发

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摘要

Large sequence polymorphisms (LSPs) or regions of differences (RDs) are molecular epidemiological and evolutionary markers used to classify Myco-bacterium tuberculosis (MTB) into East Asian (Beijing), Indo-Oceanic (IO), Euro-American (EuA) and East African Indian (EAI) lineages. The most used method is separate PCR and sequencing for each RD. We developed a single-tube multiplex PCR using four primer pairs specific to the four MTB lineages and a primer pair for species-specific RD9 with genomic DNA extracted from isolated colonies. The single-tube multiplex PCR produced lineage-specific amplicon patterns capable of differentiating the four MTB lineages. Sensitivity and specificity of the assay were 100% when differentiating MTB lineages from other species and strains of bacteria. The limit of detection of genomic MTB DNA was 12.5 ng. This single-tube multiplex PCR method offers a simple, rapid and reliable method for classification of MTB lineages based on LSPs.
机译：大序列多态性（LSPs）或差异区域（RDs）是分子流行病学和进化标记，用于将结核分枝杆菌（MTB）分为东亚（北京），印度洋（IO），欧美（EuA）和东非印度裔（EAI）血统。最常用的方法是对每个RD分别进行PCR和测序。我们开发了一种单管多重PCR，使用了四个MTB谱系特有的四个引物对和物种特异性RD9的引物对以及从分离菌落中提取的基因组DNA。单管多重PCR产生的谱系特异性扩增子模式能够区分四个MTB谱系。当将MTB谱系与其他物种和细菌菌株区分开来时，测定的灵敏度和特异性为100％。基因组MTB DNA的检出限为12.5 ng。这种单管多重PCR方法为基于LSP的MTB谱系分类提供了一种简单，快速和可靠的方法。

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