DETECTION OF Mycobacterium tuberculosis COMPLEX BY PCR-RESTRICTION FRAGMENT LENGTH POLYMORFISM ANALYSIS OF THE HSP65 GENE

Authors

  • Eduardo Eustáquio de Souza Figueiredo Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro, Brasil
  • Marlei Gomes da Silva Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro, Brasil
  • Leila de Souza Fonseca Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro, Brasil
  • Joab Trajano Silva Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro, Brasil
  • Vânia Margaret Flosi Paschoalin Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro, Brasil

Keywords:

Tecnologia de alimentos

Abstract

Mycobacterium bovis is a member of the M. tuberculosis complex, a group composed by species with high genetic homology. The pathogen is the etiological agent of bovine tuberculosis, an important zoonosis that is mainly transmitted by inhalation of infectious droplet nuclei or by ingestion of milk and crude milk derivative products from tuberculosis cows. The definitive identification of M. bovis, up to species level, is time consuming and difficult. In this work, the objective was to standardize a polymerase chain reaction followed by an enzyme restriction analysis in order to identify the M. tuberculosis complex in milk, without a microbiological isolation step. Reference strains and raw milk seeded with M. Bovis, were used as the starting material.  A 441pb fragment of the hsp65 gene was amplified and digested by two restriction enzymes BstEII and HaeIII. The obtained profile was used to identify the M. tuberculosis complex in milk. The minimum limit of detection of M. bovis in milk was 10CFU/mL. PRA methodology proved to be a specific and sensible method. It can be used to assist the microbiological and biochemical methods commonly used to identifying the bacilli in clinical samples, as milk 

Key word: Detection limit (PRA), Mycobacterium tuberculosis complex, milk Mycobacterium bovis, Restriction Enzyme Analysis (PCR),

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Published

2008-12-21

How to Cite

FIGUEIREDO, E. E. de S.; SILVA, M. G. da; FONSECA, L. de S.; SILVA, J. T.; PASCHOALIN, V. M. F. DETECTION OF Mycobacterium tuberculosis COMPLEX BY PCR-RESTRICTION FRAGMENT LENGTH POLYMORFISM ANALYSIS OF THE HSP65 GENE. Brazilian Animal Science/ Ciência Animal Brasileira, Goiânia, v. 9, n. 4, p. 1023–1033, 2008. Disponível em: https://revistas.ufg.br/vet/article/view/1381. Acesso em: 12 apr. 2025.

Issue

Vol. 9 No. 4 (2008)

Section

Veterinary Medicine

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Copyright (c) 2008 Brazilian Animal Science/ Ciência Animal Brasileira

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