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Journal of Clinical Microbiology, April 2001, p. 1559-1565, Vol. 39, No. 4
Unité de la Tuberculose et des
Mycobactéries, Institut Pasteur, F-97165 Pointe-à-Pitre
Cedex, Guadeloupe,1 and Department of
Hygiene and Microbiology, "G. D'Alessandro" University, I-90127
Palermo,2 and Department of Public
Health, University of Florence, I-50134
Florence,3 Italy
Received 27 November 2000/Returned for modification 20 January
2001/Accepted 3 February 2001
In a previous study, we proposed to associate spoligotyping and
typing with the variable number of tandem DNA repeats (VNTR) as an
alternative strategy to IS6110-restriction fragment length polymorphism (RFLP) for molecular epidemiological studies on
tuberculosis. The aim of the present study was to further evaluate this
PCR-based typing strategy and to describe the population structure of
Mycobacterium tuberculosis in another insular setting,
Sicily. A collection of 106 DNA samples from M. tuberculosis patient isolates was characterized by spoligotyping
and VNTR typing. All isolates were independently genotyped by the
standard IS6110-RFLP method, and clustering results between
the three methods were compared. The totals for the clustered isolates
were, respectively, 15, 60, and 82% by IS6110-RFLP,
spoligotyping, and VNTR typing. The most frequent spoligotype included
type 42 that missed spacers 21 to 24 and spacers 33 to 36 and derived types 33, 213, and 273 that, together represented as much as 26% of
all isolates, whereas the Haarlem clade of strains (types 47 and 50, VNTR allele 32333) accounted for 9% of the total strains. The
combination of spoligotyping and VNTR typing results reduced the number
of clusters to 43% but remained superior to the level of
IS6110-RFLP clustering (ca. 15%). All but one
IS6110-defined cluster were identified by the combination
of spoligotyping and VNTR clustering results, whereas 9 of 15 spoligotyping-defined clusters could be further subdivided by
IS6110-RFLP. Reinterpretation of previous
IS6110-RFLP results in the light of spoligotyping-VNTR typing results allowed us to detect an additional cluster that was
previously missed. Although less discriminative than
IS6110-RFLP, our results suggest that the use of the
combination of spoligotyping and VNTR typing is a good screening
strategy for detecting epidemiological links for the study of
tuberculosis epidemiology at the molecular level.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.4.1559-1565.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Genetic Diversity of Mycobacterium
tuberculosis in Sicily Based on Spoligotyping and Variable Number
of Tandem DNA Repeats and Comparison with a Spoligotyping Database
for Population-Based Analysis
*
Corresponding author. Mailing address: Unité de
la Tuberculose et des Mycobactéries, Institut Pasteur de
Guadeloupe, Morne Jolivière, BP 484, F-97165 Pointe-à-Pitre
Cedex, Guadeloupe. Phone: 590-897-665. Fax: 590-893-880. E-mail:
csola{at}pasteur.gp.
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