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首页> 外文期刊>Journal of Clinical Microbiology >Molecular Fingerprinting of Clostridium difficile Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length Polymorphism
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Molecular Fingerprinting of Clostridium difficile Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length Polymorphism

机译：艰难梭菌分离物的分子指纹图谱：脉冲场凝胶电泳与扩增的片段长度多态性

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摘要

Two molecular fingerprinting techniques, pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP), were used to investigate the epidemiological relatedness among Clostridium difficile isolates from suspected outbreaks in three general hospitals. Analysis by PFGE yielded inconclusive data as a result of extensive DNA degradation. Although this degradation could be prevented to a certain extent by the inclusion of thiourea in the electrophoresis buffer, the weak DNA banding patterns obtained in this way were still far from optimal. AFLP data were obtained by using fluorescently labeled PCR primers and analysis on an ABI PRISM automated DNA analysis platform. AFLP analysis yielded high resolution and highly reproducible DNA fingerprinting patterns from which the epidemiological relatedness among the isolates could easily be determined. AFLP results could be readily obtained within 24 h, whereas 3 to 4 days were routinely required to complete the lengthy PFGE protocol. AFLP clearly proved to be a much more fail-safe fingerprinting method for C. difficile isolates, especially for those isolates for which a standard PFGE procedure yielded inconclusive results due to DNA degradation.
机译：三种分子指纹技术分别是脉冲场凝胶电泳（PFGE）和扩增片段长度多态性（AFLP），用于调查三所综合医院疑似疫情的艰难梭菌分离株的流行病学相关性。由于DNA大量降解，PFGE分析得出的数据尚无定论。尽管可以通过在电泳缓冲液中加入硫脲在一定程度上防止这种降解，但是以这种方式获得的弱DNA谱带图谱仍远非最佳。 AFLP数据是通过使用荧光标记的PCR引物并在ABI PRISM自动化DNA分析平台上进行分析获得的。 AFLP分析产生了高分辨率和高度可重复的DNA指纹图谱，从中可以轻松确定分离株之间的流行病学相关性。 AFLP结果可在24小时内轻松获得，而通常需要3至4天才能完成冗长的PFGE方案。事实证明，AFLP是 C的一种更加故障安全的指纹识别方法。艰难梭菌分离株，特别是对于那些由于DNA降解而采用标准PFGE程序无法得出结论的分离株。

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