Progression toward an improved DNA amplification-based typing technique in the study of Mycobacterium tuberculosis epidemiology
Progression toward an improved DNA amplification-based typing technique in the study of Mycobacterium tuberculosis epidemiology
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DOI：
10.1128/jcm.01428-05
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发表时间：
2006-07-01
影响因子：
9.4
通讯作者：
Drobniewski, Francis A.
中科院分区：
文献类型：
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作者：
Gopaul, Krishna K.;Brown, Timothy J.;Drobniewski, Francis A.
虽然基于高拷贝数IS 6110的限制性片段长度多态性（HCN-RFLP）是大多数结核分枝杆菌菌株分型的金标准，但培养所需的时间和低通量使得对大量分离株进行大规模前瞻性分型不切实际。一种新方法的发展，分枝杆菌散布重复单位（MIRU），原来的可变数目串联重复（VNTR）技术的变化，可能提供一个可行的替代方案。使用基于原始12个位点MIRU（12 MIRU）、12 MIRU和剩余ETR位点的组合（15 MIRU-VNTR）以及具有额外10个新区域的扩展组（25 VNTR）的组来研究具有不同程度流行病学数据的三个人群。MIRU辨别力随着样本组大小和spoligotyping的增加而增加。结合这两种技术，使小组的大小从25个减少到14个基因座，而没有显着损失的歧视。然而，25 VNTR单独或结合spoligotyping仍然具有较低的歧视比RFLP高拷贝数的菌株。
While high-copy-number IS6110-based restriction fragment length polymorphism (HCN-RFLP) is the gold standard for typing most Mycobacterium tuberculosis strains, the time taken for culturing and low throughput make it impractical for large-scale prospective typing of large numbers of isolates. The development of a new method, mycobacterial interspersed repetitive units (MIRU), a variation of the original variable-number tandem repeat (VNTR) technique, may provide a viable alternative. Panels based on the original 12-loci MIRU (12MIRU), a combination of 12MIRU and remaining ETR loci (15MIRU-VNTR), and an extended panel with an additional 10 novel regions (25VNTR) were used to study three populations with varying degrees of epidemiological data. MIRU discrimination increased with panel size and the addition of spoligotyping. Combining these two techniques enabled a reduction in the panel size from 25 to 14 loci without a significant loss in discrimination. However, 25VNTR alone or in combination with spoligotyping still possessed weaker discrimination than RFLP for high-copy-number isolates.