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Journal of Clinical Microbiology, July 2001, p. 2453-2457, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2453-2457.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Comparison of Variable Number Tandem Repeat and IS6110-Restriction Fragment Length Polymorphism Analyses for Discrimination of High- and Low-Copy-Number IS6110 Mycobacterium tuberculosis Isolates

Rachael E. L. Barlow,1 Deborah M. Gascoyne-Binzi,1,2,* Stephen H. Gillespie,3 Anne Dickens,3 Shabnam Qamer,1 and Peter M. Hawkey1,2

The Division of Microbiology, School of Biochemistry & Molecular Biology, University of Leeds, Leeds LS2 9JT,1 Department of Microbiology, The General Infirmary, Leeds LS1 3EX,2 and Department of Medical Microbiology, Royal Free & University College Medical School, University College London, Royal Free Campus, London NW3 2PE,3 United Kingdom

Received 16 October 2000/Returned for modification 24 January 2001/Accepted 29 April 2001

The present study was designed to evaluate the use of variable number tandem repeat (VNTR) and IS6110-restriction fragment length polymorphism (RFLP) analyses in combination as a two-step strategy for discrimination (as measured by the Hunter-Gaston Discrimination Index [HGDI]) of both high- and low-copy-number IS6110 Mycobacterium tuberculosis isolates compared to IS6110-RFLP alone with an unselected collection of isolates. Individually, IS6110-RFLP fingerprinting produced six clusters that accounted for 69% of the low-copy-number IS6110 isolates (five clusters) and 5% of the high-copy-number IS6110 isolates (one cluster). A total of 39% of all the isolates were clustered (HGDI = 0.97). VNTR analysis generated a total of 35 different VNTR allele profile sets from 93 isolates (HGDI = 0.938). Combining IS6110-RFLP analysis with VNTR analysis reduced the overall percentage of clustered isolates to 29% (HGDI = 0.988) and discriminated a further 27% of low-copy-number isolates that would have been clustered by IS6110-RFLP alone. The use of VNTR analysis as an initial typing strategy facilitates further analysis by IS6110-RFLP, and more importantly, VNTR analysis subdivides some IS6110-RFLP-defined clusters containing low- and single-copy IS6110 isolates.

* Corresponding author. Mailing address: The Division of Microbiology, Department of Microbiology, The General Infirmary, Great George Street, Leeds LS1 3EX, United Kingdom. Phone: (44)-113-2335592. Fax: (44)-113-2335638. E-mail: deborahg{at}pathology.leeds.ac.uk.

Journal of Clinical Microbiology, July 2001, p. 2453-2457, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2453-2457.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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