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对从南加利福尼亚州分离出的鸟分枝杆菌复合群菌株进行的扩增片段长度多态性分析。

Amplified fragment length polymorphism analysis of Mycobacterium avium complex isolates recovered from southern California.

作者信息

Pfaller Stacy L, Aronson Timothy W, Holtzman Alan E, Covert Terry C

机构信息

National Exposure Research Laboratory, US Environmental Protection Agency, Cincinnati, OH, USA.

Education and Research Institute, Olive View - University of California Los Angeles (UCLA), Medical Center, Sylmar, Los Angeles, CA, USA.

出版信息

J Med Microbiol. 2007 Sep;56(Pt 9):1152-1160. doi: 10.1099/jmm.0.47075-0.

Abstract

Fine-scale genotyping methods are necessary in order to identify possible sources of human exposure to opportunistic pathogens belonging to the Mycobacterium avium complex (MAC). In this study, amplified fragment length polymorphism (AFLP) analysis was evaluated for fingerprinting 159 patient and environmental MAC isolates from southern California. AFLP analysis accurately identified strains belonging to M. avium and Mycobacterium intracellulare and differentiated between strains within each species. The method was also able to differentiate strains that were presumed to be genetically identical in two previous studies using large RFLP analysis with PFGE, or PCR-amplification of DNA segments located between insertion sequences IS1245 and IS1311. For M. avium, drinking-water isolates clustered more closely with each other than with patient or food isolates. Patient isolates were more genetically diverse. None of the environmental isolates shared identical AFLP patterns with patient isolates for either species. There were, however, environmental isolates that shared identical patterns, and patient isolates that shared identical patterns. A subset of the isolates, which are referred to as MX isolates due to their ambiguous identification with the Gen-Probe system, produced AFLP patterns similar to those obtained from M. intracellulare isolates. Sequence analysis of 16S rDNA obtained from the MX isolates suggests that they are strains of M. intracellulare that were not correctly identified by the M. intracellulare AccuProbe from Gen-Probe.

摘要

为了确定人类接触鸟分枝杆菌复合群(MAC)中机会性病原体的可能来源,精细基因分型方法是必要的。在本研究中,对来自南加州的159株患者和环境MAC分离株进行了扩增片段长度多态性(AFLP)分析以进行指纹识别。AFLP分析准确地鉴定了鸟分枝杆菌和胞内分枝杆菌的菌株,并区分了每个物种内的菌株。该方法还能够区分在之前两项研究中被认为基因相同的菌株,这两项研究分别使用了基于PFGE的大型RFLP分析,或对插入序列IS1245和IS1311之间的DNA片段进行PCR扩增。对于鸟分枝杆菌,饮用水分离株彼此之间的聚类比与患者或食物分离株的聚类更紧密。患者分离株的遗传多样性更高。两种物种的环境分离株与患者分离株均未共享相同的AFLP模式。然而,存在共享相同模式的环境分离株,以及共享相同模式的患者分离株。一部分分离株,由于其用Gen-Probe系统鉴定不明确而被称为MX分离株,产生了与从胞内分枝杆菌分离株获得的AFLP模式相似的模式。从MX分离株获得的16S rDNA序列分析表明,它们是胞内分枝杆菌的菌株,但未被Gen-Probe的胞内分枝杆菌AccuProbe正确鉴定。

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