First Worldwide Proficiency Study on Variable-Number Tandem-Repeat Typing of Mycobacterium tuberculosis Complex Strains
First Worldwide Proficiency Study on Variable-Number Tandem-Repeat Typing of Mycobacterium tuberculosis Complex Strains
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DOI：
10.1128/jcm.00607-11
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发表时间：
2012-03-01
影响因子：
9.4
通讯作者：
van Soolingen, Dick
中科院分区：
文献类型：
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作者：
de Beer, Jessica L.;Kremer, Kristin;van Soolingen, Dick
尽管可变数量串联重复 (VNTR) 分型已被公认为结核分枝杆菌复合体 (MTBC) 分离株 DNA 指纹分析的新标准，但外部质量控制程序尚未开发。因此，我们组织了首个24位点VNTR分型的多中心能力研究。 MTBC 菌株的 30 个 DNA 组（包括 10 个重复 DNA 样本）分布在全球 30 个不同国家的 37 个参与实验室中。 24 个实验室使用内部调整的方法，通过凝胶电泳或自动 DNA 分析仪进行片段大小测定，9 个实验室使用市售试剂盒，4 个实验室使用其他方法。 VNTR 分型的实验室内和实验室间重现性从 0% 到 100% 不等，平均值分别为 72% 和 60%。 37 个实验室中有 20 个未能扩增特定的 VNTR 基因座；如果忽略这些缺失的结果，具有 100% 实验室间重现性的实验室数量从 1 个增加到 5 个。使用商业试剂盒进行 VNTR 分型的平均实验室间重现性 (88%) 优于使用 DNA 分析仪 (70%) 或凝胶电泳 (50%) 的内部改进方法。 11 个使用内部适应的手动打字或自动打字的实验室的实验室间和实验室内再现性达到 80% 或更高，这表明这些方法可以以可靠的方式使用。总之，这项第一项多中心研究记录了 MTBC 菌株 VNTR 分型的全球质量，并强调了国际质量控制对未来改善基因分型的重要性。
Although variable-number tandem-repeat (VNTR) typing has gained recognition as the new standard for the DNA fingerprinting of Mycobacterium tuberculosis complex (MTBC) isolates, external quality control programs have not yet been developed. Therefore, we organized the first multicenter proficiency study on 24-locus VNTR typing. Sets of 30 DNAs of MTBC strains, including 10 duplicate DNA samples, were distributed among 37 participating laboratories in 30 different countries worldwide. Twenty-four laboratories used an in-house-adapted method with fragment sizing by gel electrophoresis or an automated DNA analyzer, nine laboratories used a commercially available kit, and four laboratories used other methods. The intra-and interlaboratory reproducibilities of VNTR typing varied from 0% to 100%, with averages of 72% and 60%, respectively. Twenty of the 37 laboratories failed to amplify particular VNTR loci; if these missing results were ignored, the number of laboratories with 100% interlaboratory reproducibility increased from 1 to 5. The average interlaboratory reproducibility of VNTR typing using a commercial kit was better (88%) than that of in-house-adapted methods using a DNA analyzer (70%) or gel electrophoresis (50%). Eleven laboratories using in-house-adapted manual typing or automated typing scored inter-and intralaboratory reproducibilities of 80% or higher, which suggests that these approaches can be used in a reliable way. In conclusion, this first multicenter study has documented the worldwide quality of VNTR typing of MTBC strains and highlights the importance of international quality control to improve genotyping in the future.