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Vol. 11. Issue 6.
Pages 533-556 (November - December 2005)
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Vol. 11. Issue 6.
Pages 533-556 (November - December 2005)
Original Article
Open Access
Spoligtyping e polimorfismo do gene pncA: Um cenário em duas etapas para o diagnóstico de Mycobacterium bovis em Portugal
Spoligotyping and pncA polymorphisme: A two step scenario for Mycobacterium bovis diagnosis in Portugal
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Suzana David
,
Corresponding author
suzanadavid@ihmt.unl.pt

Correspondência/Correspondence to: Suzana David, UEI de Micobacteriologia, Instituto de Higiene e Medicina Tropical. Rua da Junqueira, 96; 1349-008 Lisboa, Portugal Tel.: +351-21-3652600; Fax: +351-21-3632105.
, Clara Portugal**, Abílio Antunes***, Ana Calado*, Ângela Cardoso*, Vanessa Barros*, Luísa Sancho**, José Germano de Sousa**
* Unidade de Ensino e Investigação (UEI) de Micobacteriologia, Instituto de Higiene e Medicina Tropical (IHMT)/Universidade Nova de Lisboa (UNL), Lisboa/ Microbacteriology Teaching and Research Unit (TRU), Hygiene and Tropical Medicine Institute (HTMI)/ Nova University Lisbon (NUL), Lisbon.
** Serviço de Patologia Clínica, Hospital Fernando Fonseca, Amadora/Clinical Pathology Department, Fernando Fonseca Hospital, Amadora.
*** UEI de Clínica e Doenças Tropicais/IHMT/UNL, Lisboa/TRU of the Tropical Diseases Clinic, HTMI, NUL, Lisbon.
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Resumo

O diagnóstico diferencial do Mycobacterium bovis é importante no controlo da transmissão para a população humana e também no tratamento da doença, visto que o M. bovis é naturalmente resistente à pirazinamida. Foram analisados onze isolados clínicos do Hospital Fernando Fonseca, com um padrão de Spoligotype indicativo de M. bovis pela ausência dos espaçadores 39-43, mas que também contaram com a ausência do espaçador 38. Para a identificação destas estirpes procedeu-se à análise fenotípica da resistência à pirazinamida e estudo dos polimorfismos dos genes pncA e gyrB. Pelo estudo do polimorfismo do pncA, verificámos que as estirpes analisadas não continham a mutação específica do M. bovis. Em relação aos polimorfismos do gyrB, utilizando o kit GenoType® MTBC, as estirpes foram identificadas como pertencentes ao grupo M. tuberculosis, M. africanum subtipo II e M. canetti. O presente trabalho de investigação permitiu-nos definir novos genótipos sobre os quais deverão apoiar-se estudos bacteriológicos futuros. Entre estes foi desde já considerado importante o estudo do polimorfismo do gene pncA, devido às implicações de utilidade prática e imediata para o clínico. Avaliar a transmissão e definir grupos de risco é um objectivo para o qual se considera importante o apoio dos serviços veterinários.

Rev Port Pneumol 2005; XI (6): 533-556

Palavras-chave:
Complexo Mycobacterium tuberculosis
Mycobacterium bovis
Spoligotyping
kit GenoType® MTBC
pncA
gyrB
pirazinamida
Abstract

The differential diagnosis of Mycobacterium bovis is important in the control of transmission to the human population and also for treatment since M. bovis is naturally resistant to pyrazinamide. Eleven clinical isolates from the Fernando Fonseca Hospital with Spoligotypes indicative of M. bovis, through the absence of spacers 39-43 but that also counted with the absence of spacer 38, were analyzed. For the identification of these strains, the phenotypic analysis of pyrazinamide resistance and study of the polymorphisms of the pncA and gyrB genes were carried out. The study of the pncA polymorphism revealed that the strains analyzed did not contain the M. bovis specific mutation. In relation the gyrB polymorphisms, using the GenoType® MTBC kit, the strains were identified as belonging to the group M. tuberculosis, M. africanum subtipo II e M. canetti. The present investigation enabled us to define new genotypes on which future bacteriological studies should be based. Amongst these the study of the pncA polymorphism was considered important due to the immediate practical implications for the clinician. Evaluating transmission and defining groups of risk is an objective for which support from the veterinary services is considered relevant.

Rev Port Pneumol 2005; XI (6): 533-556

Key-words:
Mycobacterium tuberculosis complex
Mycobacterium bovis
Spoligotyping
kit GenoType® MTBC
pncA
gyrB
pyrazinamide
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