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Journal of Clinical Microbiology, March 2000, p. 1121-1126, Vol. 38, No. 3
Molecular Biology Unit, Virus Reference
Division, Central Public Health Laboratory, London NW9 5HT, United
Kingdom
Received 27 September 1999/Returned for modification 7 November
1999/Accepted 11 December 1999
The whole-genome fingerprinting technique, fluorescent
amplified-fragment length polymorphism (FAFLP) analysis, was applied to
Mycobacterium tuberculosis. Sixty-five clinical isolates
were analyzed to determine the value of FAFLP as a stand-alone
genotyping technique and to compare it with the well-established
IS6110 typing system. The genome sequence of M. tuberculosis strain H37Rv (S. T. Cole et al., Nature
393:537-544, 1998) was used to model computer-generated informative
primer combination(s), and the precision and reproducibility of FAFLP
were evaluated by comparing the results of in vitro and computer-generated experiments. Multiplex FAFLP was used to increase resolving power in a predictable and systematic fashion. FAFLP analysis
was broadly congruent with IS6110 typing for those strains with multiple IS6110 copies. It was also able to resolve an
epidemiologically unlinked group of strains with only one copy of
IS6110; up to 10% of clinical isolates may fall into this
category. For certain epidemiological investigations, it was concluded
that a combination of FAFLP and IS6110 typing would give
higher resolution than would either alone. FAFLP data were digital,
precise, reproducible, and suitable for rapid electronic dissemination,
manipulation, interlaboratory comparison, and storage in national or
international epidemiological databases. Because FAFLP samples and
analyzes base substitution across the genome as a whole, FAFLP could
generate new information about the microevolution of the M. tuberculosis complex.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Genome-Sequence-Based Fluorescent
Amplified-Fragment Length Polymorphism Analysis of
Mycobacterium tuberculosis
*
Corresponding author. Mailing address: Molecular
Biology Unit, Virus Reference Division, Central Public Health
Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone:
181 200 4400. Fax: 181 200 1569. E-mail:
carnold{at}hgmp.mrc.ac.uk.
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