First worldwide proficiency study on variable-number tandem-repeat typing of Mycobacterium tuberculosis complex strains

Jessica L. De Beer, Kristin Kremer, Csaba Ködmön, Philip Supply, Dick Van Soolingen, David Alexander, Ewa Augustynowicz-Kopeć, Urska Bidovec-Stojkovic, Martin J. Boeree, Timothy Brown, Daniela M. Cirillo, Laura Cruz, Susana David, Raul Diaz, Horng Yunn Dou, Jason T. Evans, Maryse Fauville-Dufaux, Margaret M. Fitzgibbon, Darío García De Viedma, Maria GlobanRamona Groenheit, Marjo Haanperä-Heikkinen, Alexander Indra, Kai Man Kam, Rebecca Kramer, Shang Mei, Stefan Niemann, Mihaela Obrovac, Erik M. Rasmussen, Guislaine Refrégier, Jaime A. Robledo, Sofia Samper, Meenu K. Sharma, Wladimir Sougakoff, Petras Stakenas, Ruth Stavrum, Philip N. Suffys, Juraj Trenkler, Takayuki Wada, Robin M. Warren

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)662-669
Number of pages8
JournalJournal of Clinical Microbiology
Volume50
Issue number3
DOIs
Publication statusPublished - Mar 2012

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ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

De Beer, J. L., Kremer, K., Ködmön, C., Supply, P., Van Soolingen, D., Alexander, D., Augustynowicz-Kopeć, E., Bidovec-Stojkovic, U., Boeree, M. J., Brown, T., Cirillo, D. M., Cruz, L., David, S., Diaz, R., Dou, H. Y., Evans, J. T., Fauville-Dufaux, M., Fitzgibbon, M. M., García De Viedma, D., ... Warren, R. M. (2012). First worldwide proficiency study on variable-number tandem-repeat typing of Mycobacterium tuberculosis complex strains. Journal of Clinical Microbiology, 50(3), 662-669. https://doi.org/10.1128/JCM.00607-11