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

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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.

Original languageEnglish
JournalMemorias do Instituto Oswaldo Cruz
Issue number6
Pages (from-to)396-403
Number of pages8
Publication statusPublished - 2017
Externally publishedYes


  • Antibiotics, Antitubercular/pharmacology
  • DNA, Bacterial/genetics
  • Drug Resistance, Multiple, Bacterial/drug effects
  • Genotype
  • Humans
  • Isoniazid/pharmacology
  • Microbial Sensitivity Tests
  • Multiplex Polymerase Chain Reaction
  • Mycobacterium tuberculosis/drug effects
  • Phenotype
  • Rifampin/pharmacology
  • Tuberculosis, Multidrug-Resistant/microbiology


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