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

doi:10.1590/0074-02760160376

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

Research output: Contribution to journal › A1: Web of Science-article › peer-review

14 Downloads (Pure)

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.

Original language	English
Journal	Memorias do Instituto Oswaldo Cruz
Volume	112
Issue number	6
Pages (from-to)	396-403
Number of pages	8
ISSN	0074-0276
DOIs	https://doi.org/10.1590/0074-02760160376
Publication status	Published - 2017
Externally published	Yes

Keywords

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

Access to Document

10.1590/0074-02760160376

2017mioc0396

Cite this

Santos, P. F. G. D., Costa, E. R. D., Ramalho, D. M., Rossetti, M. L., Barcellos, R. B., Nunes, L. D. S., Esteves, L. S., Rodenbusch, R., Anthony, R. M., Bergval, I., Sengstake, S., Viveiros, M., Kritski, A., & Oliveira, M. M. (2017). Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus. Memorias do Instituto Oswaldo Cruz, 112(6), 396-403. https://doi.org/10.1590/0074-02760160376

@article{06083a94cfd84597bbc000005a720afa,

title = "Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype{\textregistered}MTBDRplus",

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{\textregistered} 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{\textregistered}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{\textregistered}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{\textregistered}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{\textregistered}MTBDRplus, MLPA showed similar sensitivity to detect INH and RIF resistance. The results obtained by the MLPA and Genotype{\textregistered}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.",

keywords = "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",

author = "Santos, {Paula Fernanda Gon{\c c}alves Dos} and Costa, {Elis Regina Dalla} and Ramalho, {Daniela M} and Rossetti, {Maria Lucia} and Barcellos, {Regina Bones} and Nunes, {Luciana de Souza} and Esteves, {Leonardo Souza} and Rodrigo Rodenbusch and Anthony, {Richard M} and Indra Bergval and Sarah Sengstake and Miguel Viveiros and Afr{\^a}nio Kritski and Oliveira, {Martha M}",

note = "FTX",

year = "2017",

doi = "10.1590/0074-02760160376",

language = "English",

volume = "112",

pages = "396--403",

journal = "Memorias do Instituto Oswaldo Cruz",

issn = "0074-0276",

publisher = "Fundaco Oswaldo Cruz",

number = "6",

}

Santos, PFGD, Costa, ERD, Ramalho, DM, Rossetti, ML, Barcellos, RB, Nunes, LDS, Esteves, LS, Rodenbusch, R, Anthony, RM, Bergval, I, Sengstake, S, Viveiros, M, Kritski, A & Oliveira, MM 2017, 'Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus', Memorias do Instituto Oswaldo Cruz, vol. 112, no. 6, pp. 396-403. https://doi.org/10.1590/0074-02760160376

Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus. / Santos, Paula Fernanda Gonçalves Dos; Costa, Elis Regina Dalla; Ramalho, Daniela M et al.
In: Memorias do Instituto Oswaldo Cruz, Vol. 112, No. 6, 2017, p. 396-403.

Research output: Contribution to journal › A1: Web of Science-article › peer-review

TY - JOUR

T1 - Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus

AU - Santos, Paula Fernanda Gonçalves Dos

AU - Costa, Elis Regina Dalla

AU - Ramalho, Daniela M

AU - Rossetti, Maria Lucia

AU - Barcellos, Regina Bones

AU - Nunes, Luciana de Souza

AU - Esteves, Leonardo Souza

AU - Rodenbusch, Rodrigo

AU - Anthony, Richard M

AU - Bergval, Indra

AU - Sengstake, Sarah

AU - Viveiros, Miguel

AU - Kritski, Afrânio

AU - Oliveira, Martha M

N1 - FTX

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

KW - Antibiotics, Antitubercular/pharmacology

KW - DNA, Bacterial/genetics

KW - Drug Resistance, Multiple, Bacterial/drug effects

KW - Genotype

KW - Humans

KW - Isoniazid/pharmacology

KW - Microbial Sensitivity Tests

KW - Multiplex Polymerase Chain Reaction

KW - Mycobacterium tuberculosis/drug effects

KW - Phenotype

KW - Rifampin/pharmacology

KW - Tuberculosis, Multidrug-Resistant/microbiology

U2 - 10.1590/0074-02760160376

DO - 10.1590/0074-02760160376

M3 - A1: Web of Science-article

C2 - 28591399

SN - 0074-0276

VL - 112

SP - 396

EP - 403

JO - Memorias do Instituto Oswaldo Cruz

JF - Memorias do Instituto Oswaldo Cruz

IS - 6

ER -

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

Abstract

Keywords

Access to Document

Fingerprint

Cite this