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Molecular approach to differential identification of Mycobacterium species and detection of drug-resistant tuberculosis.

机译：鉴别结核分枝杆菌种类和检测耐药结核的分子方法。

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摘要

The recent increase in infections caused by tuberculous, as well as non-tuberculous Mycobacterium, has intensified the requirement for a rapid and reliable molecular identification tool as an alternative to current conventional methods. The differentiation between Mycobacterium tuberculosis and Mycobacterium bovis, along with the rapid detection of the resistance to first-line antituberculosis drug, pyrazinamide, are of great importance for the control of tuberculosis. The hypothesis of this study was that sequence diversity at specific targets of the Mycobacterium DNA can provide for rapid and accurate differential identification of mycobacterial species and detection of antimicrobial resistance. A unique sequence-based identification algorithm called "MycoAlign" that utilized the hypervariable sequence of the internal transcribed spacer (ITS) region of the rDNA gene was created for identification of Mycobacterium species. The study proved the feasibility of the MycoAlign assay as an alternative tool to the conventional identification methods. Sequence analysis of tested isolates has also showed five previously uncharacterized Mycobacterium species from five different immunocompromised patients with a general clinical picture of pulmonary infection. An evaluation of the phenotypic properties, the mycolic acid-HPLC profile, and the 16S rDNA-dependant phylogenetic analysis of the organism has placed the new species within the slow-growing Mycobacterium species and supported the conclusion that the described organism represented a new slow-growing Mycobacterium species, for which the name Mycobacterium nebraskiae sp. nov. was proposed. A temperature mediated Heteroduplex analysis (TMHA) assay using the denaturing high performance liquid chromatography (DHPLC) was also developed using the pncA gene as genetic target for rapid detection of pyrazinamide resistance and differentiation between M. tuberculosis and M. bovis. Chromatographic patterns generated by the two reference probes provided for the rapid detection of PZA-R with the simultaneous ability to distinguish between M. tuberculosis and M. bovis . In conclusion, the two described assays represented an alternative molecular tool that provided for rapid and accurate differential identification of all clinically important mycobacterial species including M. tuberculosis and M. bovis, and the detection of the pyrazinamide resistance among tuberculosis causing agents.
机译：由结核病和非结核分枝杆菌引起的感染的最近增加，使得对快速和可靠的分子鉴定工具的需求增加，以替代当前的常规方法。结核分枝杆菌和牛分枝杆菌之间的区别，以及对一线抗结核药物吡嗪酰胺的耐药性的快速检测，对于控制结核病至关重要。这项研究的假设是，分枝杆菌DNA特定靶点的序列多样性可以提供快速，准确的鉴别分枝杆菌种类和检测抗菌素耐药性的方法。创建了一种独特的基于序列的识别算法，称为“ MycoAlign”，该算法利用了rDNA基因内部转录间隔区（ITS）区域的高变序列来鉴定分枝杆菌。这项研究证明了MycoAlign分析作为常规鉴定方法的替代工具的可行性。对测试菌株的序列分析还显示了来自五名不同免疫功能低下患者的五种以前未鉴定的分枝杆菌，具有肺部感染的一般临床表现。对表型特性，霉菌酸-HPLC谱图以及依赖于该生物的16S rDNA进行系统发育分析的评估已将新物种置于缓慢生长的分枝杆菌物种中，并支持了以下结论：所描述的生物代表了一种新的慢细菌。生长的分枝杆菌物种，其名称为“ Nebraskiae sp。”。十一月被提出。还开发了使用变性高效液相色谱（DHPLC）的温度介导的异源双链分析（TMHA）方法，该方法使用pncA基因作为遗传靶标，用于快速检测吡嗪酰胺抗性以及结核分枝杆菌和牛分枝杆菌之间的分化。由两种参考探针产生的色谱图谱，可用于快速检测PZA-R，同时具有区分结核分枝杆菌和牛分枝杆菌的能力。总之，上述两种测定方法代表了一种替代的分子工具，可快速准确地鉴别包括结核分枝杆菌和牛分枝杆菌在内的所有临床上重要的分枝杆菌菌种，并检测引起结核病的菌种对吡嗪酰胺的抗性。

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  • 作者

    Mohamed, Amr Mohamed Abdel Fattah.;

  • 作者单位

    University of Nebraska Medical Center.;

  • 授予单位 University of Nebraska Medical Center.;
  • 学科 Biology Microbiology.;Health Sciences Public Health.;Health Sciences Pathology.
  • 学位 Ph.D.
  • 年度 2004
  • 页码 172 p.
  • 总页数 172
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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