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Journal of Clinical Microbiology, November 2002, p. 4191-4196, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.4191-4196.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Novel Allelic Variants of Mycobacteria Isolated in Brazil as Determined by PCR-Restriction Enzyme Analysis of hsp65

A. da Silva Rocha,1 A. M. Werneck Barreto,2 C. E. Dias Campos,2 M. Villas-Bôas da Silva,2 L. Fonseca,3 M. H. Saad,4 W. M. Degrave,1 and P. N. Suffys1*

Biochemistry and Molecular Biology Departmentand Leprosy Laboratory, Oswaldo Cruz Institute, Oswaldo Cruz Foundation,1 Reference Center Prof. Hélio Fragaand,2 Federal University of Rio de Janeiro,3 Rio de Janeiro, Brazil4

Received 9 April 2002/ Returned for modification 22 July 2002/ Accepted 8 August 2002

Human isolates of Mycobacterium collected in 16 different states of Brazil were submitted to PCR-restriction analysis (PRA) of a 439-bp fragment of the hsp65 gene with HaeIII and BstEII. Fourteen allelic variants not described in clinical isolates so far were observed among 36 (10%) of 356 Brazilian strains, including a new pattern for Mycobacterium scrofulaceum, M. intracellulare, and M. flavescens, two new patterns for M. fortuitum, three new patterns each for M. gordonae and M. terrae, and one new pattern for M. avium complex-like strains. Two unidentified strains each also presented a new pattern, strongly suggesting that Mycobacterium genotypes are distributed biogeographically. The PRA procedure was also performed with 43 reference isolates belonging to 34 species, adding a further six new patterns to the identification algorithm. A database containing the normalized restriction patterns of both enzymes was constructed. Patterns available on the Internet can be introduced into this database, which will make possible the comparison of genotypes from isolates from different parts of the world.

* Corresponding author. Mailing address: Laboratory of Molecular Biology and Diagnosis of Infectious Diseases, DBBM, IOC, Fiocruz, Av. Brasil 4365, Manguinhos 21045-900, Rio de Janeiro, Brazil. Phone: 55-21-2598 4289. Fax: 55-21-2270 9997. E-mail: psuffys{at}ioc.fiocruz.br.

Journal of Clinical Microbiology, November 2002, p. 4191-4196, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.4191-4196.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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