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Memorias do Instituto Oswaldo Cruz
Jun, 2017;112 (6): 396-403.

Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus.

Paula Fernanda Gonçalves Dos Santos, Elis Regina Dalla Costa, Daniela M Ramalho, Maria Lucia Rossetti, Regina Bones Barcellos, Luciana de Souza Nunes, Leonardo Souza Esteves, Rodrigo Rodenbusch, Richard M Anthony, Indra Bergval, Sarah Sengstake, Miguel Viveiros, Afrânio Kritski, Martha M Oliveira
Author Information
  1. Paula Fernanda Gonçalves Dos Santos: Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Programa Acadêmico de Tuberculose, Programa de Pós-Graduação em Clínica Médica, Rio de Janeiro, RJ, Brasil.
  2. Elis Regina Dalla Costa: Fundação Estadual de Produção e Pesquisa em Saúde, Centro de Desenvolvimento Científico e Tecnológico, Porto Alegre, RS, Brasil.
  3. Daniela M Ramalho: Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Programa Acadêmico de Tuberculose, Programa de Pós-Graduação em Clínica Médica, Rio de Janeiro, RJ, Brasil.
  4. Maria Lucia Rossetti: Fundação Estadual de Produção e Pesquisa em Saúde, Centro de Desenvolvimento Científico e Tecnológico, Porto Alegre, RS, Brasil.
  5. Regina Bones Barcellos: Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Programa Acadêmico de Tuberculose, Programa de Pós-Graduação em Clínica Médica, Rio de Janeiro, RJ, Brasil.
  6. Luciana de Souza Nunes: Universidade Federal do Rio Grande do Sul, Centro de Biotecnologia, Programa de Pós-Graduação em Biologia Celular e Molecular, Porto Alegre, RS, Brasil.
  7. Leonardo Souza Esteves: Fundação Estadual de Produção e Pesquisa em Saúde, Centro de Desenvolvimento Científico e Tecnológico, Porto Alegre, RS, Brasil.
  8. Rodrigo Rodenbusch: Fundação Estadual de Produção e Pesquisa em Saúde, Centro de Desenvolvimento Científico e Tecnológico, Porto Alegre, RS, Brasil.
  9. Richard M Anthony: Royal Tropical Institute, KIT Biomedical Research, Amsterdam, The Netherlands.
  10. Indra Bergval: Royal Tropical Institute, KIT Biomedical Research, Amsterdam, The Netherlands.
  11. Sarah Sengstake: Royal Tropical Institute, KIT Biomedical Research, Amsterdam, The Netherlands.
  12. Miguel Viveiros: Universidade Nova de Lisboa, Instituto de Higiene e Medicina Tropical, Unidade de Microbiologia Médica, Global Health and Tropical Medicine, Lisboa, Portugal.
  13. Afrânio Kritski: Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Programa Acadêmico de Tuberculose, Programa de Pós-Graduação em Clínica Médica, Rio de Janeiro, RJ, Brasil.
  14. Martha M Oliveira: Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Programa Acadêmico de Tuberculose, Programa de Pós-Graduação em Clínica Médica, Rio de Janeiro, RJ, Brasil.
PMID: 28591399 DOI: 10.1590/0074-02760160376

Abstract

BACKGROUND: To cope with the emergence of multidrug-resistant tuberculosis (MDR-TB), new molecular methods that can routinely be used to screen for a wide range of drug resistance related genetic markers in the Mycobacterium tuberculosis genome are urgently needed.
OBJECTIVE: To evaluate the performance of multiplex ligaton-dependent probe amplification (MLPA) against Genotype® MTBDRplus to detect resistance to isoniazid (INHr) and rifampicin (RIFr).
METHOD: 96 culture isolates characterised for identification, drug susceptibility testing (DST) and sequencing of rpoB, katG, and inhA genes were evaluated by the MLPA and Genotype®MTBDRplus assays.
RESULTS: With sequencing as a reference standard, sensitivity (SE) to detect INHr was 92.8% and 85.7%, and specificity (SP) was 100% and 97.5%, for MLPA and Genotype®MTBDRplus, respectively. In relation to RIFr, SE was 87.5% and 100%, and SP was 100% and 98.8%, respectively. Kappa value was identical between Genotype®MTBDRplus and MLPA compared with the standard DST and sequencing for detection of INHr [0.83 (0.75-0.91)] and RIFr [0.93 (0.88-0.98)].
CONCLUSION: Compared to Genotype®MTBDRplus, MLPA showed similar sensitivity to detect INH and RIF resistance. The results obtained by the MLPA and Genotype®MTBDRplus assays indicate that both molecular tests can be used for the rapid detection of drug-resistant TB with high accuracy. MLPA has the added value of providing information on the circulating M. tuberculosis lineages.

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MeSH Term

Antibiotics, Antitubercular
DNA, Bacterial
Drug Resistance, Multiple, Bacterial
Genotype
Humans
Isoniazid
Microbial Sensitivity Tests
Multiplex Polymerase Chain Reaction
Mycobacterium tuberculosis
Phenotype
Rifampin
Tuberculosis, Multidrug-Resistant

Chemicals

Antibiotics, Antitubercular
DNA, Bacterial
Isoniazid
Rifampin
Journal Article
Article has an altmetric score of 1

Word Cloud

Created with Highcharts 10.0.0MLPAGenotype®MTBDRplustuberculosisdrugresistancedetectINHrRIFrsequencing100%molecularcanusedMycobacteriumDSTassaysstandardsensitivitySE8%SP5%respectively98valuedetection[00]BACKGROUND:copeemergencemultidrug-resistantMDR-TBnewmethodsroutinelyscreenwiderangerelatedgeneticmarkersgenomeurgentlyneededOBJECTIVE:evaluateperformancemultiplexligaton-dependentprobeamplificationGenotype®MTBDRplusisoniazidrifampicinMETHOD:96cultureisolatescharacterisedidentificationsusceptibilitytestingrpoBkatGinhAgenesevaluatedRESULTS:reference92857%specificity97relation87Kappaidenticalcompared8375-0919388-0CONCLUSION:ComparedshowedsimilarINHRIFresultsobtainedindicatetestsrapiddrug-resistantTBhighaccuracyaddedprovidinginformationcirculatingMlineagesDetectionresistance:comparisonassayversus

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