Mycobacterium tuberculosis complex CRISPR genotyping: improving efficiency, throughput and discriminative power of 'spoligotyping' with new spacers and a microbead-based hybridization assay
Mycobacterium tuberculosis complex CRISPR genotyping: improving efficiency, throughput and discriminative power of 'spoligotyping' with new spacers and a microbead-based hybridization assay
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DOI：
10.1099/jmm.0.016949-0
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发表时间：
2010-03-01
影响因子：
3
通讯作者：
Sola, Christophe
中科院分区：
文献类型：
--
作者：
Zhang, Jian;Abadia, Edgar;Sola, Christophe
本研究的目的是实施一种基于微珠的“spoligotyping”技术，并通过增加一组25个额外的间隔区来评估改进，我们希望这些间隔区能提供对主要遗传组1（PGG 1）菌株的更高分辨率。我们证实了使用43间隔板的经典技术的高灵敏度和再现性，并且我们获得了基于膜的技术和基于微珠的技术之间的完美一致。我们进一步证明了用于区分PGG 1临床分离株的扩展的68个间隔区格式的区分能力的增加，特别是对于东非-印度进化枝。最后，我们定义了一个有限的，但信息量很大的减少10个间隔区面板集，可以提供一个更具成本效益的选择，在资源有限的国家实施，并可以减少需要额外的VNTR（可变数目的串联重复序列）基因分型工作的分子流行病学研究。我们还提出了一个经济分析比较膜为基础的和微珠为基础的技术。
The aims of the present study were to implement a microbead-based 'spoligotyping' technique and to evaluate improvements by the addition of a panel of 25 extra spacers that we expected to provide an increased resolution on principal genetic group 1 (PGG 1) strains. We confirmed the high sensitivity and reproducibility of the classical technique using the 43 spacer panel and we obtained perfect agreement between the membrane-based and the microbead-based techniques. We further demonstrated an increase in the discriminative power of an extended 68 spacer format for differentiation of PGG 1 clinical isolates, in particular for the East African-Indian clade. Finally, we define a limited yet highly informative reduced 10 spacer panel set which could offer a more cost-effective option for implementation in resource-limited countries and that could decrease the need for additional VNTR (variable number of tandem repeats) genotyping work in molecular epidemiological studies. We also present an economic analysis comparing membrane-based and microbead-based techniques.