title: Genetic diversity of the Mycobacterium tuberculosis Beijing family in Brazil and Mozambique and relation with infectivity and induction of necrosis in THP-1 cells.
abstract: BACKGROUND
  The success of the Mycobacterium tuberculosis Beijing (MtbB) lineage in different geographical regions has been attributed to high transmission, increased virulence, drug resistance and rapid adaptation to the host. In some countries of secondary MtbB dispersion like South Africa and Peru, rising prevalence of the Beijing strains is registered. However, in neighboring countries to affected regions such as Mozambique and Brazil, respectively, the prevalence of these strains is still low and this could be due to biological particularities of the circulating MtbB strains and/or differentiated host susceptibility.
  
  
  OBJECTIVE
  To characterize genetically and phenotypically MtbB strains isolated in Brazil (n = 8) and Mozambique (n = 17).
  
  
  METHODS
  This is a descriptive study of genotypes of the MtbB isolates, determined by spoligotyping, MIRU-VNTR typing, analysis of the IS6110 copy number in the NTF region and screening for mutations in mutT2, mutT4, rpoB, katG and pks 15/1 genes. Virulence-associated properties of the studied isolates were verified in the in vitro model of infection of human THP-1 cells.
  
  
  RESULTS
  The genotypes defined by the 24VNTRs were distinct for all isolates included in this study and presented an HGDI of 0.997. The VNTR patterns with seven copies of MIRU26 and seven copies of QUB26, representative for the previously described MtbB genotype B0, dominant in Russia, were detected in 38.5% of the studied isolates. In addition, all isolates presented RD105 deletion and a 7 bp insertion in pks15/1 gene. Almost all tested strains belonged to the RD181 sublineage, with the exception of two strains from Mozambique of RD150 sublineage. Combined analysis of the NTF region integrity and mutations in mutT genes showed that 62.5% and 47% of isolates obtained in Brazil and Mozambique, respectively, were of the ancestral genotype. The virulence index of the ancient isolates, evaluated in the THP-1 cells, was significantly lower than that of the modern genotype group.
  
  
  CONCLUSIONS
  These data demonstrate genotype particularities of the Beijing strains isolated in Brazil and Mozambique, two countries of low prevalence of the MtbB lineage in local Mtb populations. In contrast to the neighboring countries with high prevalence of the MtbB strains of modern sublineage, significant proportions of the isolates obtained in Brazil and Mozambique were presented by the strains of the ancient sublineage. Our data suggest that lower virulence of the ancient strains, compared with the modern strains, could be involved in the slow spread of the MtbB strains in some regions.
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Lia Lima
authors.initials: LL
authors.lastname: Gomes
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Sidra Ezidrio Gonçalves
authors.initials: SE
authors.lastname: Vasconcellos
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Harrison Magdinier
authors.initials: HM
authors.lastname: Gomes
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Atina Ribeiro
authors.initials: AR
authors.lastname: Elias
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Adalgiza
authors.initials: A
authors.lastname: da Silva Rocha
authors.affiliation: Laboratory of Biology of Recognition, Universidade Estadual do Norte Fluminense, Rio de Janeiro, Brazil.
authors.forename: Simone C M
authors.initials: SC
authors.lastname: Ribeiro
authors.affiliation: Laboratory of Bacterial Pathogenesis and Molecular Biology, Unicamp, São Paulo, Brazil.
authors.forename: Alessandra Costa
authors.initials: AC
authors.lastname: Panunto
authors.affiliation: Instituto Adolfo Lutz, São Paulo, Brazil.
authors.forename: Lucilaine
authors.initials: L
authors.lastname: Ferrazoli
authors.affiliation: Instituto Adolfo Lutz, São Paulo, Brazil.
authors.forename: Maria Alice
authors.initials: MA
authors.lastname: da Silva Telles
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Araujo Marelo Emanuel
authors.initials: AM
authors.lastname: Ivens de
authors.affiliation: Laboratorio de Micobacteriologia Molecular do Centro de Pesquisas em Doenças Infecciosas e Parasitarias, University Hospital Clementino Fraga Filho, UFRJ, Rio de Janeiro, Brazil.
authors.forename: Afranio Lineu
authors.initials: AL
authors.lastname: Kritski
authors.affiliation: Laboratory of Molecular Microbiology, St. Petersburg Pasteur Institute, St. Petersburg, Russia.
authors.forename: Igor
authors.initials: I
authors.lastname: Mokrousov
authors.affiliation: Laboratory of Microbiology, Research Institute of Phthisiopulmonology, St. Petersburg, Russia.
authors.forename: Olga A
authors.initials: OA
authors.lastname: Manicheva
authors.affiliation: Laboratory of Biology of Recognition, Universidade Estadual do Norte Fluminense, Rio de Janeiro, Brazil.
authors.forename: Elena
authors.initials: E
authors.lastname: Lasunskaia
authors.affiliation: Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil. Electronic address: psuffys@gmail.com.
authors.forename: Philip Noel
authors.initials: PN
authors.lastname: Suffys
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D000068476:Beijing; D001938:Brazil / Q000453:epidemiology; D002478:Cells, Cultured; D004269:DNA, Bacterial / Q000235:genetics; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D009073:Mozambique / Q000453:epidemiology; D009154:Mutation / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000472:pathogenicity; D009336:Necrosis / Q000382:microbiology; D014376:Tuberculosis / Q000453:epidemiology / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial
keywords: Beijing; Genotypes; Mozambique and Brazil; Mycobacterium tuberculosis; Virulence