Gladys Lopez-Avalos1, Guadalupe Gonzalez-Palomar1, Martín Lopez-Rodriguez2, Carlos Arturo Vazquez-Chacon3, Gustavo Mora-Aguilera4, Juan Antonio Gonzalez-Barrios5, Juan Carlos Villanueva-Arias6, Manuel Sandoval-Diaz6, Ulises Miranda-Hernández1, Ikuri Alvarez-Maya7. 1. Medical and Pharmaceutical Biotechnology, The Center for Research and Applied Technology in Jalisco (CIATEJ), Av. Normalistas No. 800, Col. Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco, Mexico. 2. Public Health Laboratory of Jalisco, Guadalajara, Jalisco, Mexico. 3. Institute for Epidemiologic Diagnosis and Reference (InDRE), México City, Mexico. 4. Phytopathology, Postgraduates College, Campus Montecillo, Texcoco, Mexico. 5. Laboratory of Genomic Medicine, Regional Hospital October 1, ISSSTE, Avenida Instituto Politécnico Nacional No. 1669, 07760 México, DF, Mexico. 6. Health Minister of Jalisco, Calle Dr. Baeza Alzaga No. 107 Zona Centro, C.P. 44100, Guadalajara, Jalisco, Mexico. 7. Medical and Pharmaceutical Biotechnology, The Center for Research and Applied Technology in Jalisco (CIATEJ), Av. Normalistas No. 800, Col. Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco, Mexico. Electronic address: ialvarez@ciatej.mx.
Abstract
OBJECTIVES: The objectives of this study were to analyse the frequency of gene mutations associated with antitubercular drug resistance in clinical samples from the population of Jalisco State (Mexico) and to evaluate the genetic variability of Mycobacterium tuberculosis and multidrug-resistant (MDR) M. tuberculosis strains to describe the frequency of various families. METHODS: Clinical isolates of M. tuberculosis obtained from Jalisco State were analysed. Isolates were subjected to drug susceptibility testing, and mutations were characterised by sequencing, followed by genotyping using spoligotyping and mycobacterial interspersed repetitive units-variable-number of tandem repeats (MIRU-VNTR). Moreover, the prevalence of mutations was analysed by phylogenetic lineages. RESULTS: Resistant strains were analysed by sequencing of katG, inhA and rpoB genes to determine the presence of mutations associated with isoniazid and rifampicin resistance. In MDR, monoresistant and polyresistant isolates, mutations were found in 17 (54.84%) of 31 strains. Spoligotyping identified six different strain lineages [T1 (25.40%), H3 (7.94%), MANU (4.76%), X1 (3.17%), EAI5 (1.59%) and LAM1 (1.59%)], with the remaining strains identified as orphans. In additional tree-based identification, a dendrogram of spoligotype patterns generated five different similarity clusters. When combining 24-loci MIRU-VNTR and spoligotyping approaches, the results shows that there is no cluster formation, indicating low transmission of the samples. CONCLUSIONS: This study using spoligotyping and MIRU-VNTR showed that the analysed strains were not related to each other since no two identical strains were found. Families with the highest prevalence in the study were orphans followed by T family.
OBJECTIVES: The objectives of this study were to analyse the frequency of gene mutations associated with antitubercular drug resistance in clinical samples from the population of Jalisco State (Mexico) and to evaluate the genetic variability of Mycobacterium tuberculosis and multidrug-resistant (MDR) M. tuberculosis strains to describe the frequency of various families. METHODS:Clinical isolates of M. tuberculosis obtained from Jalisco State were analysed. Isolates were subjected to drug susceptibility testing, and mutations were characterised by sequencing, followed by genotyping using spoligotyping and mycobacterial interspersed repetitive units-variable-number of tandem repeats (MIRU-VNTR). Moreover, the prevalence of mutations was analysed by phylogenetic lineages. RESULTS: Resistant strains were analysed by sequencing of katG, inhA and rpoB genes to determine the presence of mutations associated with isoniazid and rifampicin resistance. In MDR, monoresistant and polyresistant isolates, mutations were found in 17 (54.84%) of 31 strains. Spoligotyping identified six different strain lineages [T1 (25.40%), H3 (7.94%), MANU (4.76%), X1 (3.17%), EAI5 (1.59%) and LAM1 (1.59%)], with the remaining strains identified as orphans. In additional tree-based identification, a dendrogram of spoligotype patterns generated five different similarity clusters. When combining 24-loci MIRU-VNTR and spoligotyping approaches, the results shows that there is no cluster formation, indicating low transmission of the samples. CONCLUSIONS: This study using spoligotyping and MIRU-VNTR showed that the analysed strains were not related to each other since no two identical strains were found. Families with the highest prevalence in the study were orphans followed by T family.
Authors: Graziele Lima Bello; Franciele Costa Leite Morais; Sheile Pinheiro de Jesus; Jonas Michel Wolf; Mirela Gehlen; Isabela Neves de Almeida; Lida Jouca de Assis Figueiredo; Tainá Dos Santos Soares; Regina Bones Barcellos; Elis Regina Dalla Costa; Silvana Spíndola de Miranda; Maria Lucia Rosa Rossetti Journal: Mem Inst Oswaldo Cruz Date: 2020-04-17 Impact factor: 2.743