Marie Sylvianne Rabodoarivelo1, A Brandao2, M C Cergole Novella3, A G C Bombonatte4, B Imperiale5, N Rakotosamimanana6, N Morcillo7, V Rasolofo1, J C Palomino8, A Martin9. 1. Mycobacteria Unit, Institut Pasteur de ; Ecole Doctorale Sciences de la Vie et de l'Environnement, Université d'Antananarivo, Antananarivo, Madagascar. 2. Instituto Adolfo Lutz Núcleo de Tuberculose e Micobacteriosis Sao Paulo, Sao Paulo; Fundação Oswaldo Cruz, Rio de Janeiro, Brazil. 3. Instituto Adolfo Lutz, CRL-Santo Andre, Brazil. 4. Instituto Adolfo Lutz, CRL-Santos, Brazil. 5. Institute of Experimental Medicine-CONICET, National Academy of Medicine, Buenos Aires, Argentina. 6. Mycobacteria Unit, Institut Pasteur de , Antananarivo, Madagascar. 7. Dr. Cetrangolo Hospital, Reference Laboratory of Tuberculosis Control Program, Buenos Aires, Argentina. 8. Laboratory of Microbiology, Department of Biochemistry and Microbiology, Ghent University, Gent, Belgium. 9. Université catholique de Louvain (UCL), Institute of Experimental and Clinical Research (IREC), Laboratory of Medical Microbiology (MBLG), Brussels, Belgium.
Abstract
Background: In low-income countries, rapid detection of tuberculosis (TB) drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF) and isoniazid (INH) resistance. Methods: This was a multicenter study. Molecular detection of RIF and INH resistance was performed directly from smear-positive TB sputa spotted on a slide, FTA card, GenoCard, and ethanol using the Genotype MTBDRplus assay. The performance of the DNA extraction method from each storage support to detect drug resistance was assessed by calculating their sensitivity and specificity compared to the phenotypic method. Results: From all sites, the overall sensitivity and specificity for RIF-resistance detection was 88% and 85%, respectively, for slides, 86% and 92%, respectively, for GenoCard, 87% and 89%, respectively, for FTA card, and 88% and 92%, respectively, for ethanol. For INH-resistance detection, the overall sensitivity and specificity was 82% and 90%, respectively, for slides, 85% and 96%, respectively, for GenoCard, 86% and 92%, respectively, for FTA card, and 86% and 94%, respectively, for ethanol. Conclusion: Smear slides and filter cards showed to be very useful tools to facilitate DNA extraction from sputum samples with the potential to accelerate the detection of drug resistance in remote areas.
Background: In low-income countries, rapid detection of tuberculosis (TB) drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF) and isoniazid (INH) resistance. Methods: This was a multicenter study. Molecular detection of RIF and INH resistance was performed directly from smear-positive TB sputa spotted on a slide, FTA card, GenoCard, and ethanol using the Genotype MTBDRplus assay. The performance of the DNA extraction method from each storage support to detect drug resistance was assessed by calculating their sensitivity and specificity compared to the phenotypic method. Results: From all sites, the overall sensitivity and specificity for RIF-resistance detection was 88% and 85%, respectively, for slides, 86% and 92%, respectively, for GenoCard, 87% and 89%, respectively, for FTA card, and 88% and 92%, respectively, for ethanol. For INH-resistance detection, the overall sensitivity and specificity was 82% and 90%, respectively, for slides, 85% and 96%, respectively, for GenoCard, 86% and 92%, respectively, for FTA card, and 86% and 94%, respectively, for ethanol. Conclusion: Smear slides and filter cards showed to be very useful tools to facilitate DNA extraction from sputum samples with the potential to accelerate the detection of drug resistance in remote areas.
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