Characterization of mutations in clinical isolates of Mycobacterium tuberculosis resistant to 1st and 2nd line drugs and the relationship between mutations and phenotypic resistance levels to isoniazid.

• Grant number: 17/16082-7
• Support Opportunities: Regular Research Grants
• Start date: November 01, 2017
• End date: April 30, 2020
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Rosangela Siqueira de Oliveira
• Grantee: Rosangela Siqueira de Oliveira
• Host Institution: Instituto Adolfo Lutz (IAL). Coordenadoria de Controle de Doenças (CCD). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Ana Angélica Bulcão Portela Lindoso ; Angela Pires Brandao ; Cristina Viana Niero ; Juliana Maira Watanabe Pinhata ; Lucilaine Ferrazoli

Abstract

Tuberculosis (TB) is an infectious disease of universal distribution, constituting a serious public health problem in Brazil and in the world. Isoniazid is one of the main anti-TB drugs, being used both in the treatment of active and latent disease. Mutations in several genes have been found in isoniazid-resistant strains of Mycobacterium tuberculosis (MTB). Depending on the mutated gene, M. tuberculosis may express different levels of phenotypic resistance. Serum concentration in patients treated with isoniazid may reach 5 ¼g/ml. In the susceptibility testing, the concentration tested is 0.1 ¼g/mL, which has no relationship with serum concentrations. Once the in vitro resistance to isoniazid is detected, it is replaced by streptomycin, an injectable and more toxic drug to the patient. MTB resistant to rifampicin and isoniazid characterizes multidrug-resistance (MDR)-TB and treatment can last from 18 to 24 months. Treatment failure of MDR-TB may progress to extended-resistance TB (TB-XDR), in which the MTB, besides being MDR, is resistant to any fluoroquinolone and to at least one of the second-line injectable drugs. DNA sequencing of MTB isolates is a rapid molecular method for evaluating resistance to anti-TB drugs, detecting known as well as new mutations. This project aims to (i) standardize a quantitative susceptibility testing to isoniazid in more clinically significant doses and to analyze mutations in genes related to resistance to this drug; (ii) evaluate the evolution of isoniazid resistance during the treatment; (iii) describe the frequency of MDR and XDR strains of MTB using DNA sequencing; (iv) standardize the MALDI-TOF technique for anti-TB drugs resistance detection, and (v) describe treatment outcomes of patients whose isolates were included in this study. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

WATANABE PINHATA, JULIANA MAIRA; BRANDAO, ANGELA PIRES; MENDES, FLAVIA DE FREITAS; DA SILVA RABELLO, MICHELLE CHRISTIANE; FERRAZOLI, LUCILAINE; DE OLIVEIRA, ROSANGELA SIQUEIRA. Correlating genetic mutations with isoniazid phenotypic levels of resistance in Mycobacterium tuberculosis isolates from patients with drug-resistant tuberculosis in a high burden setting. EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, v. 40, n. 12, JUL 2021. (12/51756-5, 17/16082-7)

FOWLER, PHILIP W.; BARILAR, IVAN; BATTAGLIA, SIMONE; BORRONI, EMANUELE; BRANDAO, ANGELA PIRES; BRANKIN, ALICE; CABIBBE, ANDREA MAURIZIO; CARTER, JOSHUA; CIRILLO, DANIELA MARIA; CLAXTON, PAULINE; et al. Epidemiological cut-off values for a 96-well broth microdilution plate for high-throughput research antibiotic susceptibility testing of M. tuberculosis. European Respiratory Journal, v. 60, n. 4, p. 22-pg., 2022-10-01. (17/16082-7, 12/51756-5)

[ANONYMOUS]. A data compendium associating the genomes of 12,289 Mycobacterium tuberculosis isolates with quantitative resistance phenotypes to 13 antibiotics. PLOS BIOLOGY, v. 20, n. 8, p. 25-pg., 2022-08-01. (12/51756-5, 17/16082-7)

RODRIGUES, CAMILLA; MOORE, DAVID; CROOK, DERRICK W.; CIRILLO, DANIELA M.; FOWLER, PHILIP W.; IQBAL, ZAMIN; ISMAIL, NAZIR A.; MISTRY, NERGES; NIEMANN, STEFAN; PETO, TIM E. A.; et al. Genome-wide association studies of global Mycobacterium tuberculosis resistance to 13 antimicrobials in 10,228 genomes identify new resistance mechanisms. PLOS BIOLOGY, v. 20, n. 8, p. 27-pg., 2022-08-01. (12/51756-5, 17/16082-7)

WATANABE PINHATA, JULIANA MAIRA; BRANDAO, ANGELA PIRES; GALLO, JULIANA FAILDE; SIQUEIRA DE OLIVEIRA, ROS ANGELA; FERRAZOLI, LUCILAINE. GenoType MTBDRsl for detection of second-line drugs and ethambutol resistance in multidrug-resistant Mycobacterium tuberculosis isolates at a high-throughput laboratory. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, v. 105, n. 2, p. 9-pg., 2023-02-01. (12/51756-5, 17/16082-7)

BRANDAO, ANGELA PIRES; WATANABE PINHATA, JULIANA MAIRA; SIMONSEN, VERA; OLIVEIRA, ROSANGELA SIQUEIRA; GHISI, KELEN TEIXEIRA; SILVA RABELLO, MICHELLE CHRISTIANE; FUKASAVA, SUELY; FERRAZOLI, LUCILAINE. Transmission of Mycobacterium tuberculosis presenting unusually high discordance between genotypic and phenotypic resistance to rifampicin in an endemic tuberculosis setting. TUBERCULOSIS, v. 125, DEC 2020. (17/16082-7, 12/51756-5)

TANIA MATSUI; JULIANA MAÍRA WATANABE PINHATA; MICHELLE CHRISTIANE DA SILVA RABELLO; ANGELA PIRES BRANDÃO; LUCILAINE FERRAZOLI; SYLVIA CARDOSO LEÃO; CRISTINA VIANA-NIERO; ROSANGELA SIQUEIRA DE OLIVEIRA. Frequency of first and second-line drug resistance-associated mutations among resistant Mycobacterium tuberculosis clinical isolates from São Paulo, Brazil. Memórias do Instituto Oswaldo Cruz, v. 115, 2020-05-08. (17/16082-7)