RESUMO
AIM: Certification of V. cholerae strains stored at State collection of pathogenic microorganisms and cell cultures SCPM - Obolensk. MATERIALS AND METHODS: 50 V. cholerae strains were studied. Real-time PCR with primers to genes ctxA, ctxB, ace, zot, tcpA, toxR, hlyA, rtxC, rfbO1, ompU, ompW was used. RESULTS: Membership of the studied strains in V. cholerae species was confirmed by molecular-biological methods. 46 strains belong to O1 serogroup, 42 of those - E1 Tor toxigenic, having all the virulence genes and 4 non-toxigenic strains. A strain had ace, zot, tcpA, toxR, rtxC, hlyA, ompU genes. 2 strains had ace, toxR, rtxC, hlyA genes, a strain had only toxR gene which is a global regulatory gene. 2 of the 4 serogroup O1 strains were non-toxigenic and had all the virulence genes (ctxA, ctxB, ace, zot, tcpA, toxR, rtxC, hlyA, ompU). 1 non-toxigenic strain has ace, zot, toxR, hlyA, ompUgenes, and the other - toxR, hlyAgenes. CONCLUSION: Genome certificates of all the V. cholerae strains stored in State collection of pathogenic microorganisms and cell cultures SCPM - Obolensk were created. Markers of epidemic threat - ctxA, ctxB, tcpA, toxR and additional virulence genes were determined.
Assuntos
Proteínas de Fímbrias/genética , Ilhas Genômicas/genética , Vibrio cholerae , Virulência/genética , Toxina da Cólera/genética , Primers do DNA , Genes Reguladores , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição/genética , Vibrio cholerae/classificação , Vibrio cholerae/genética , Vibrio cholerae/patogenicidadeRESUMO
Description of the pyrazinamide-resistant clinical strains of M. tuberculosis derived from sputum of patients treated in TB clinics in Tula was made (June, 2001 - July, 2002). It was demonstrated that 30.3% (n = 91) strains were resistant to pyrazinamide. It was found out that these strains were resistant to other antituberculosis drugs in most cases. The method of PCR-sequencing was used to find the mutations in the gene pncA determining resistance to pyrazinamide. 44 different types of mutations localized in 28 codons were detected. The predominance of the mutations in 57 (13.2%), 63 (7.7%), 97 (7.7%), 12 (6.6%), 103 (6.6%) codons and in -11 (6.6%) promoter ofp ncA was observed in the pyrazinamide-resistant strains. Several new mutations determining resistance of the clinical strains of M. tuberculosis to pyrazinamide were described. A high correlation between resistance of mycobacteria to pyrazinamide and activity of pyrazinamidase was observed.
Assuntos
Amidoidrolases/genética , Antituberculosos/farmacologia , Farmacorresistência Bacteriana , Mycobacterium tuberculosis/efeitos dos fármacos , Pirazinamida/farmacologia , Humanos , Mutação , Mycobacterium tuberculosis/isolamento & purificação , Regiões Promotoras Genéticas , Escarro/microbiologiaRESUMO
The critical concentration of canamycin for the MGIT Bactec 960 system, which has been determined on a panel of the tho--roughly characterized genetically heterogeneous clinical Mycobacterium tuberculosis isolated in the Central Region of Russia, is equal to 1.25 microg/ml.