伊朗西北部的结核病传播：使用 MIRU-VNTR、ETR-VNTR 和 IS6110-RFLP 方法。

Tuberculosis transmission in Northwest of Iran: using MIRU-VNTR, ETR-VNTR and IS6110-RFLP methods.

机构信息

Infectious Diseases and Tropical Medicine Research Center, Golgasht Ave, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Infect Genet Evol. 2011 Jan;11(1):124-31. doi: 10.1016/j.meegid.2010.09.013. Epub 2010 Oct 14.

DOI:10.1016/j.meegid.2010.09.013

PMID:20951237

Abstract

Tuberculosis is the main cause for adults' death from a single infectious agent killing about 2 million people every year. Identifying the source of transmission infection is necessary to decrease tuberculosis. In this study we used IS6110-RFLP, MIRU-VNTR and ETR-VNTR methods to determine transmission dynamic of tuberculosis in Northwest of Iran from March 2004 to March 2005. 156 culture positive isolates (51 from Orumieh and 105 from Tabriz) were collected. IS6110-RFLP was done by standardized method and MIRU-VNTR was done by 12 primers specific for 12 MIRU-VNTR loci by polymerase chain reaction and ETR was carried out by primers specific for ETR-A, ETR-B and ETR-C as well. Genetic relationship analyses were performed by MVSP software and MIRU-VNTRplus website. The results revealed that the copy number of IS6110 in isolates ranged from 0 to 17. One hundred and seven unique patterns and 16 shared patterns by 2-10 isolates were determined by IS6110-RFLP, and 116 distinct patterns were identified by MIRU-VNTR including 21 clustered patterns and 95 unique patterns. Having combined MIRU-VNTR and ETR-VNTR data, we increased the discriminatory power of typing method, thus 25 clusters were identified. Loci 10, 26, 40 in MIRU and ETR-A were highly discriminatory. Distribution of lineages in our study according to tentative data in MIRU-VNTRplus database was multi-matches 9.09%, Bovis 9.74%, S 8.44%, New-1 0.65%, Uganda I 0.65%, Haarlem 0.65%, Cameroon 0.65% and X 0.65%. The minimum estimate for tuberculosis proportion due to transmission was 20.1% with IS6110-RFLP and was 18.8% with MIRU and ETR. In conclusion, tuberculosis sanitation status in northwest of Iran due to minimum estimate of tuberculosis transmission was satisfactory, and the reactivation of tuberculosis was responsible for more than 80% of new tuberculosis. According to the results and drawbacks of IS6110-RFLP method we strongly recommend MIRU-VNTRplus ETR-VNTR as an easy, reliable, reproducible and highly discriminatory power method.

摘要

结核病是导致成年人因单一感染源而死亡的主要原因，每年约有 200 万人因此死亡。确定传播感染的来源对于减少结核病至关重要。本研究于 2004 年 3 月至 2005 年 3 月期间采用 IS6110-RFLP、MIRU-VNTR 和 ETR-VNTR 方法，对伊朗西北部的结核病传播动态进行了研究。共收集了 156 例培养阳性的分离株（51 例来自乌鲁米耶，105 例来自大不里士）。采用标准化方法进行 IS6110-RFLP 分析，采用 12 对针对 12 个 MIRU-VNTR 基因座的引物进行聚合酶链反应（PCR）进行 MIRU-VNTR 分析，采用针对 ETR-A、ETR-B 和 ETR-C 的引物进行 ETR 分析。采用 MVSP 软件和 MIRU-VNTRplus 网站进行遗传关系分析。结果显示，分离株中 IS6110 的拷贝数范围为 0 至 17。通过 IS6110-RFLP 确定了 107 个独特模式和 16 个由 2-10 个分离株共享的模式，通过 MIRU-VNTR 确定了 116 个独特模式，其中包括 21 个聚类模式和 95 个独特模式。结合 MIRU-VNTR 和 ETR-VNTR 数据，我们提高了分型方法的区分能力，因此确定了 25 个聚类。MIRU 中的 10、26 和 40 个基因座和 ETR-A 具有高度的区分能力。根据 MIRU-VNTRplus 数据库中的暂定数据，本研究中的谱系分布为多匹配 9.09%、牛型分枝杆菌 9.74%、S 型 8.44%、新 1 型 0.65%、乌干达 I 型 0.65%、哈雷姆型 0.65%、喀麦隆型 0.65%和 X 型 0.65%。结核病传播导致的结核病比例最低估计值为 20.1%，采用 IS6110-RFLP 方法，为 18.8%。采用 MIRU 和 ETR。结论：根据最低估计值，伊朗西北部的结核病卫生状况因结核病传播而令人满意，而结核病的再激活占新发病例的 80%以上。根据 IS6110-RFLP 方法的优缺点，我们强烈推荐 MIRU-VNTRplus ETR-VNTR 作为一种简单、可靠、可重复和具有高度区分能力的方法。