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首页> 外文期刊>Journal of Clinical Microbiology >Genotypic detection of Mycobacterium tuberculosis rifampin resistance: comparison of single-strand conformation polymorphism and dideoxy fingerprinting.
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Genotypic detection of Mycobacterium tuberculosis rifampin resistance: comparison of single-strand conformation polymorphism and dideoxy fingerprinting.

机译：结核分枝杆菌利福平抗性的基因型检测：单链构象多态性和双脱氧指纹图谱的比较。

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摘要

Detection of mutations in the rpoB gene of Mycobacterium tuberculosis can be used as an accurate predictor of rifampin resistance in the majority of strains tested. Simple but highly accurate screening methods must be developed for the detection of these mutations. Either DNA sequence analysis or single-strand conformation polymorphism (SSCP) screening can be used to detect rpoB mutations, but these techniques either are expensive or yield results that may prove difficult to interpret when used in a clinical setting. This report describes the use of dideoxy fingerprinting (ddF) as a postamplification screening method to identify rifampin-resistant genotypes. The ddF protocol was performed on the amplified rpoB fragment with no preparatory steps, thus making ddF practical for laboratories equipped for polyacrylamide gel electrophoresis. When compared with the results of SSCP analysis, ddF results were more easily interpreted and contained more sequence-dependent information that facilitated differentiation of functionally significant and silent mutations. The ddF method was used for genotypic determination of rifampin susceptibility of 20 multidrug-resistant strains of M. tuberculosis. The results of this analysis were concordant with DNA sequence analysis and conventional clinical laboratory methods.
机译：结核分枝杆菌rpoB基因突变的检测可以用作大多数测试菌株对利福平耐药性的准确预测指标。必须开发出简单但高度准确的筛选方法来检测这些突变。 DNA序列分析或单链构象多态性（SSCP）筛选均可用于检测rpoB突变，但这些技术要么昂贵，要么产生的结果可能在临床环境中难以解释。该报告描述了使用双脱氧指纹图谱（ddF）作为扩增后筛选方法来鉴定耐利福平的基因型。 ddF规程是在扩增的rpoB片段上进行的，无需任何准备步骤，因此ddF对于配备聚丙烯酰胺凝胶电泳技术的实验室很实用。与SSCP分析的结果相比，ddF结果更容易解释，并包含更多的序列依赖性信息，有助于区分功能上重要的沉默突变。 ddF方法用于对20株结核分枝杆菌的多药耐药菌株的利福平敏感性进行基因型测定。该分析的结果与DNA序列分析和常规临床实验室方法一致。

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