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Journal of Clinical Microbiology, August 2006, p. 2851-2856, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.00705-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Rapid Detection of the Mycobacterium tuberculosis Beijing Genotype and Its Ancient and Modern Sublineages by IS6110-Based Inverse PCR

Laboratory of Molecular Microbiology, St. Petersburg Pasteur Institute, 197101 St. Petersburg,¹ Laboratory of Microbiology of Tuberculosis, The Research Institute of Phthisiopulmonology, 193063 St. Petersburg, Russia,² Public Central Laboratory, Beijing Pediatric Institute, Beijing Children's Hospital Affiliated to Capital University of Medical Sciences, Beijing 100045, People's Republic of China,³ Department of Pathogenic Bacteria, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria,⁴ National Institute of Hygiene and Epidemiology, Hanoi,⁵ Pham Ngoc Thach Tuberculosis and Lung Diseases Centre, Ho Chi Minh City, Vietnam⁶

Received 4 April 2006/ Returned for modification 11 May 2006/ Accepted 26 May 2006

The Mycobacterium tuberculosis Beijing genotype strains appear to be hypervirulent and associated with multidrug-resistant tuberculosis. Therefore, the development of a both rapid and simple method to detect the M. tuberculosis Beijing genotype is of clinical interest per se. Previously, we described a simple and fast approach to detect the Beijing genotype based on IS6110 inverse-PCR typing. Here, we evaluated this method against a large, diverse, and recent collection of strains. The study sample included 866 M. tuberculosis strains representing but not limited to the regions in Russia, Europe, and East Asia where the Beijing genotype is endemic. Based on a spoligotyping method, 408 strains were identified as Beijing genotypes; they were additionally subdivided into ancient and modern sublineages based on the analysis of the NTF locus. All strains were further subjected to the IS6110-based inverse PCR. All of the Beijing genotype strains were found to have identical two-band (ancient sublineage) or three-band (modern sublineage) profiles that were easily recognizable and distinct from the profiles of the non-Beijing strains. Therefore, we suggest using IS6110-based inverse-PCR typing for the correct identification of the Beijing genotype and its major sublineages. The method is fast and inexpensive and does not require additional experiments but instead is implemented in the routine typing method of M. tuberculosis.