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Vol. 10. Issue 3.
Pages 195-204 (May - June 2004)
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Vol. 10. Issue 3.
Pages 195-204 (May - June 2004)
ARTIGO ORIGINAL/ORIGINAL ARTICLE
Open Access
Identificação molecular pelo método de Spoligotyping de estirpes do complexo Mycobacterium tuberculosis isoladas no Hospital Fernando Fonseca
Molecular identification using Spoligotyping of strains from the Mycobacterium tuberculosis complex isolated from the Hospital Fernando Fonseca
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Suzana David*, Clara Portugal**, Abílio Antunes***, Ângela Cardoso*, Ana Calado*, Barros Vanessa*, Luísa Sancho**
* Unidade de Ensino e Investigação (UEI) de Micobacteriologia, Instituto de Higiene e Medicina Tropical (IHMT)/Universidade Nova de Lisboa (UNL).
** Laboratório de Bacteriologia, Hospital Fernando Fonseca, Amadora.
*** UEI de Clínica e Doenças/IHMT/UNL.
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RESUMO

O Spoligotyping foi utilizado na genotipagem de 219 isolados do complexo Mycobacterium tuberculosis de doentes do Hospital Fernando Fonseca. Esta técnica, baseada na metodologia PCR, analisa uma região cromossómica específica do complexo M. tuberculosis, o locus DR (Direct Repeats).

Com o auxílio de uma base de dados internacional, mostramos que 29,2 % dos Spoligotypes predominantes pertenciam à família LAM (Latino-American Mediterranean). A família LAM 9, com 12,3 %, deixou-nos atentos à possível importação da doença de populações oriundas da América do Sul, onde tem sido frequentemente detectada. Foram identificadas as famílias genotípicas T1 e Haarlem, com 6,4 % e 8,7 %, respectivamente, apresentando uma frequência do tipo Europeia. A família Beijing, representando 1,4 %, pode estar relacionada com um problema emergente no nosso país, relativo a uma recente imigração de populações asiáticas e da Europa Oriental.

Foram encontrados isolados com um Spoligotype do tipo M. bovis em elevada percentagem – 3,7 %. Na Europa, esta infecção é extremamente rara, podendo este resultado não decorrer da infecção por M. bovis, mas por M. bovis BCG (devido à vacinação ou eventuais terapias combase no BCG recombinante) ou M. africanum (face à proximidade das duas espécies).

Uma percentagem elevada dos Spoligotypes (21,4 %) não foi identificada pela base de dados.

Este estudo é o primeiro entre nós e poderá ser o ponto de partida para a criação de uma base de dados com importantes repercussões sobre o programa nacional de luta contra a tuberculose.

REV PORT PNEUMOL 2004; X (3): 195-204

Palavras-chave:
Complexo Mycobacterium tuberculosis
Spoligotyping
Portugal
ABSTRACT

Spoligotyping was used in the genotyping of 219 isolates of the Mycobacterium tuberculosis complex, from patients of the Hospital Fernando Fonseca. This technique, based on PCR methodology, analyses a region of the chromosome specific of the Mycobacterium tuberculosis complex, the DR locus (Direct Repeat).

With the aid of an international database, we showed that the predominant Spoligotypes belonged to the LAM family (Latino-AmericanMediterranean), 29.2 %. The LAM 9 family, with 12.3 %, left us attentive to the possible import of the disease through populations from South America, were it has been frequently identified. The genotypic families T1 and Haarlem, with 6.4 % and 8.7 % respectively, represented a frequency typical to Europe. The Beijing family, with 1.4 %, may represent an emerging problem in our country due to recent immigration of Asian and Eastern European populations.

Isolates with a Spoligotype of the M. bovis type were found at a high percentage, 3.7 %. In Europe, this infection is extremely rare suggesting the result may not be due to M. bovis infection but to M. bovis BCG (due to vaccination or eventual recombinant BCG based therapies), or M. africanum (due to the proximity of the two species).

A high percentage of the Spoligotypes were not identified by the database, 21.4 %.

This is the first study of this type amongst us and may be the starting point for the creation of a data base with important consequences on the national program against tuberculosis.

REV PORT PNEUMOL 2004; X (3): 195-204

Key-words:
Mycobacterium tuberculosis complex
Spoligotyping
Portugal
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BIBLIOGRAFIA
[1.]
D. Van Soolingen.
Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements.
J Intern Med, 249 (2001), pp. 1-26
[2.]
P.J. Bifani, B. Mathema, Z. Liu, S.L. Moghazeh, B. Shopsin, B. Tempalski, J. Driscoll, R. Frothingham, J.M. Musser, P. Alcabes, B.N. Kreiswirth.
Identification of a W variant outbreak of Mycobacterium tuberculosis via population-based molecular epidemiology.
JAMA, 282 (1999), pp. 2321-2327
[3.]
J.R. Glynn, J. Whiteley, P.J. Bifani, K. Kremer, D. Van Soolingen.
Worldwide occurrence of Beijing/W strains of Mycobacterium tuberculosis: a systematic review.
Emerg Infect Dis, 8 (2002), pp. 843-849
[4.]
I. Filliol, J.R. Driscoll, D. Van Soolingen, B.N. Kreiswirth, K. Kremer, G. Valetudie, D.D. Anh, R. Barlow, D. Banerjee, P.J. Bifani, K. Brudey, A. Cataldi, R.C. Cooksey, D.V. Cousins, J.W. Dale, O.A. Dellagostin, F. Drobniewski, G. Engelmann, S. Ferdinand, D. Gascoyne-Binzi, M. Gordon, M.C. Gutierrez, W.H. Haas, H. Heersma, G. Kallenius, E. Kassa-Kelembho, T. Koivula, H.M. Ly, A. Makristathis, C. Mammina, G. Martin, P. Mostrom, I. Okrousov, V. Narbonne, O. Narvskaya, A. Nastasi, S.N. Niobe-Eyangoh, J.W. Pape, V. Rasolofo-Razanamparany, M. Ridell, M.L. Rossetti, F. Stauffer, H. Takiff, J. Texier-Maugein, V. Vincent, J.H. De Waard, C. Sola, N. Rastogi.
Global distribution of Mycobacterium tuberculosis spoligotypes.
Emerg Infect Dis, 8 (2002), pp. 1347-1349
[5.]
C. Sola, I. Filliol, M.C. Gutierrez, I. Mokrousov, V. Vincent, N. Rastogi.
Spoligotype database of Mycobacterium tuberculosis: biogeographic distribution of shared types and epidemiologic and phylogenetic perspectives.
Emerg Infect Dis, 7 (2001), pp. 390-396
[7.]
World Health Organization, Health Report, 1999.
[8.]
H.L. David, T.M. Jahan, A. Jumin, J. Grandry, E.H. Lehman.
Numerical taxonomy analysis of Mycobacterium africanum.
Int J Systematic Bacteriol, (1978), pp. 467-472
[9.]
S. Niemann, E. Richter, S. Rusch-Gerdes.
among members of the Mycobacterium tuberculosis complex by molecular and biochemical features: evidence for two pyrazinamide-susceptible subtypes of M. bovi.
J Clin Microbiol, 38 (2000), pp. 152-157
[10.]
C. Viana-Niero, C. Gutierrez, C. Sola, I. Filliol, F. Boulahbal, V. Vincent, N. Rastogi.
Genetic diversity of Mycobacterium africanum clinical isolates based on IS6110-restriction fragment length polymorphism analysis, spoligotyping, and variable number of tandem DNA repeats.
J Clin Microbiol, 39 (2001), pp. 57-65
[11.]
G. Källenius, T. Koivula, S. Ghebremichael, S.E. Hoffner, R. Norberg, E. Svensson, F. Dias, B.-I. Marklund, S.B. Svenson.
Evolution and clonal traits of Mycobacterium tuberculosi complex in Guine-Bissau.
J Clin Microbiol, 37 (1999), pp. 3872-3878
[12.]
A. Rivero, M. Marquez, J. Santos, A. Pinedo, M.A. Sanchez, A. Esteve, S. Samper, C. Martin.
High rate of tuberculosis reinfection during a nosocomial outbreak of multidrug-resistant tuberculosis caused by Mycobacterium bovis strain B.
Clin Infect Dis, 32 (2001), pp. 159-161
[13.]
R.R. Kazwala, C.J. Daborn, J.M. Sharp, D.M. Kam Barage, S.F. Jiwa.
Int J Tuberc Lung Dis, 5 (2001), pp. 87-91
[14.]
J.D.A. Van Embden, M.D. Cave, J.T. Crawford, J.W. Dale, K.D. Eisenach, B. Gicquel, P. Hermans, Martín, R. McAdam, T.M. Shinnick, P.M. Small.
Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology.
J Clin Microbiol, 31 (1993), pp. 406-409
[15.]
M.J. Covas, L. Brum, I. Portugal, J.L. Bernardo, M. Viveiros, H. David, P. Ferrinho.
Estudo epidemiológico de um surto de tuberculose multirresistente na área metroplitana de Lisboa.
Revista Portuguesa de Doenças Infecciosas, 1–2 (1998), pp. 38-44
[16.]
M.M. Hannan, H. Peres, F. Maltez, A.C. Hayward, J. Machado, A. Morgado, R. Proenca, M.R. Nelson, J. Bico, D.B. Young, B.S. Gazzard.
Investigation and control of a large outbreak of multidrug resistant tuberculosis at a central Lisbon hospital.
J Hospital Infect, 47 (2001), pp. 91-97
[17.]
I. Portugal, M.J. Covas, L. Brum, M. Viveiros, P. Ferrinho, J. Moniz-Pereira, H. David.
Outbreak of multiple drug-resistant tuberculosis in Lisbon: detection by restriction fragment length polymorphism analysis.
Int J Tuberc Lung Dis, 3 (1999), pp. 207-213
[18.]
P.M.A. Groenen, A.E. Bunschoten, D. Van Soolingen, J.D.A. Van Embden.
Nature of DNA polymorphism in the direct repeat cluster of Mycobcaterium tuberculosis; Application for strain differentiation by a novel method.
Mol Microbiol, 10 (1993), pp. 1057-1065
Copyright © 2004. Sociedade Portuguesa de Pneumologia/SPP
Pulmonology
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