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Journal of Clinical Microbiology, May 2005, p. 2148-2154, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2148-2154.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Targeted Hybridization of IS6110 Fingerprints Identifies the W-Beijing Mycobacterium tuberculosis Strains among Clinical Isolates

TB Center, Public Health Research Institute, Newark, New Jersey,¹ Mycobacteria Reference Unit, Laboratory of Infectious Diseases and Perinatal Screening, National Institute of Public Health and the Environment (RIVM), The Netherlands,² INSERM U629, Institut Pasteur de Lille, Lille, France³

Received 1 November 2004/ Returned for modification 7 December 2004/ Accepted 1 January 2005

Targeted IS6110-based RFLP genotyping can be applied to rapidly identify specific groups of biomedically/epidemiologically relevant Mycobacterium tuberculosis clinical isolates. One such group is the W-Beijing strain family (also known as Beijing/W), implicated in significant nosocomial and community outbreaks worldwide. Using previously defined criteria, we developed a simple and accurate method to identify members of the W-Beijing family, based on rehybridization of Southern blot membranes used previously in routine IS6110 DNA fingerprint analysis. The hybridization probe constructed ("W-Beijing polyprobe") contains the PCR-amplified fragments specific for three M. tuberculosis chromosomal loci used for the identification of W-Beijing strains. The targets include the dnaA-dnaN and NTF regions and the direct repeat locus. A total of 526 selected clinical isolates (representative of 253 different IS6110-defined strain types) were analyzed using the W-Beijing polyprobe. A total of 148 isolates from this collection were found to be members of the W-Beijing phylogenetic lineage, comprising 106 strains from the W-Beijing family (46 clusters) and 42 related isolates. Rehybridization results were confirmed by computer-assisted analysis. The sensitivity and specificity of this method were estimated at 98.7% and 99.7%, respectively. This study demonstrates that the W-Beijing polyprobe can accurately and reliably discriminate members of the W-Beijing phylogenetic lineage and the W-Beijing family of M. tuberculosis strains.

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