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首页> 外文期刊>Journal of Clinical Microbiology >Experimental versus In Silico Fluorescent Amplified Fragment Length Polymorphism Analysis of Mycobacterium tuberculosis: Improved Typing with an Extended Fragment Range
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Experimental versus In Silico Fluorescent Amplified Fragment Length Polymorphism Analysis of Mycobacterium tuberculosis: Improved Typing with an Extended Fragment Range

机译：结核分枝杆菌的实验性与硅胶荧光扩增的片段长度多态性分析：扩展片段范围可提高打字效率

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Whole-genome fingerprinting fluorescent amplified fragment length polymorphism (FAFLP) data were compared with in silico data for the sequenced strains of Mycobacterium tuberculosis (H37Rv and CDC1551). For this G+C-rich genome, many predicted fragments were not detected experimentally. For H37Rv, only 108 (66%) of the 163 predicted EcoRI-MseI fragments between 100 and 500 bp were visualized in vitro. FAFLP was also used to identify polymorphism in 10 clinical isolates of M. tuberculosis characterized previously by IS6110 typing, examining fragments of up to 1,000 bp in size rather than up to 500 bp as was done previously. Five isolates had unique IS6110 profiles and were not known to be epidemiologically related, two isolates were the same single-band IS6110 type but were not known to be epidemiologically related, and the remaining three isolates were epidemiologically related with identical IS6110 profiles. Analysis of fragments in the 500- to 1,000-bp range using nonselective primers differentiated better between strains than analysis of fragments in the 50- to 500-bp range using a set of four selective primers. Seventeen polymorphic fragments were identified between 500 and 1,000 bp in size compared with nine polymorphic fragments between 50 and 500 bp. Using the 500- to 1,000-bp analysis, a level of discrimination similar to that of IS6110 typing was achieved which, unlike the IS6110 typing, was able to differentiate the two M. tuberculosis strains, each of which had only a single copy of IS6110.
机译：将全基因组指纹荧光扩增片段长度多态性（FAFLP）数据与结核分枝杆菌（H37Rv和CDC1551）测序菌株的计算机数据进行了计算机比较。对于这个富含G + C的基因组，实验上没有检测到许多预测的片段。对于H37Rv，在100到500 bp之间的163个预测的 Eco RI- Mse I片段中，只有108个（66％）在体外可见。 FAFLP还被用于鉴定10个 M临床分离株的多态性。以前以IS 6110 分型为特征的结核病，检查的片段最大为1000 bp，而不是以前的最大500 bp。 5株分离株具有独特的IS 6110 谱，并且不具有流行病学相关性;两个分离株具有相同的单带IS 6110 类型，但未知与流行病学相关，其余三个分离株在流行病学上具有相同的IS 6110 谱。与使用一组四个选择性引物对50至500 bp范围内的片段进行分析相比，使用非选择性引物对500至1,000 bp范围内的片段进行分析的菌株之间的区别更好。在500至1,000 bp之间鉴定出17个多态性片段，而在50至500 bp之间则有9个多态性片段。使用500到1,000 bp的分析，可以达到与IS 6110 类型相似的辨别水平，与IS 6110 类型不同，它能够区分两个 M。结核菌株，每个菌株只有一个IS 6110 拷贝。

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