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Optimizing Tuberculosis Testing for Basic Laboratories

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  • Laboratory of Infectious Diseases Research, Department of Microbiology, Faculty of Sciences, Universidad Peruana Cayetano Heredia, Lima, Peru; Asociación Benefica Prisma, Lima, Peru; Department of International Health, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland; Wellcome Trust Centre for Clinical Tropical Medicine, Department of Infectious Diseases and Immunity, Imperial College London Hammersmith Hospital Campus, London, United Kingdom

Optimal tuberculosis testing usually involves sputum centrifugation followed by broth culture. However, centrifuges are biohazardous and scarce in the resource-limited settings where most tuberculosis occurs. To optimize tuberculosis testing for these settings, centrifugation of 111 decontaminated sputum samples was compared with syringe-aspiration through polycarbonate membrane-filters that were then cultured in broth. To reduce the workload of repeated microscopic screening of broth cultures for tuberculosis growth, the colorimetric redox indicator 2,3-diphenyl-5-(2-thienyl) tetrazolium chloride was added to the broth, which enabled naked-eye detection of culture positivity. This combination of filtration and colorimetric growth-detection gave similar results to sputum centrifugation followed by culture microscopy regarding mean colony counts (43 versus 48; P = 0.6), contamination rates (0.9% versus 1.8%; P = 0.3), and sensitivity (94% versus 95%; P = 0.7), suggesting equivalency of the two methods. By obviating centrifugation and repeated microscopic screening of cultures, this approach may constitute a more appropriate technology for rapid and sensitive tuberculosis diagnosis in basic laboratories.

Author Notes

*Address correspondence to Carlton A. Evans, Department of Microbiologia (LID), Universidad Peruana Cayetano Heredia, Av. Honorio Delgado #430, Urb. Ingeniería, San Martin de Porres, Lima, Peru. E-mail: Carlton.Evans@IFHAD.org†These authors contributed equally to this article.

Financial support: This study and/or members of the research team were funded by Innovation for Health and Development, the Foundation for Innovative New Diagnostics, the Wellcome Trust, the Sir Halley Stewart Trust, the Civil Society Challenge Fund of the Department for International Development of the British Government, and the Bill and Melinda Gates Foundation.

Authors' addresses: Eric Ramos, Beatriz Herrera, Willi Quino, Jessica Alvarado, Louis Grandjean, Laura Martin, and Jonathan M. Sherman, Laboratory of Infectious Diseases Research, Department of Microbiology, Faculty of Sciences, Universidad Peruana Cayetano Heredia, Lima, Peru. Samuel G. Schumacher and Rosario Montoya, Laboratory of Infectious Diseases Research, Department of Microbiology, Faculty of Sciences, Universidad Peruana Cayetano Heredia, Lima, Peru and Asociación Benefica Prisma, Lima, Peru. Mark Siedner, Department of International Health, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Room W5515, Baltimore, MD 21205. Robert H. Gilman, Laboratory of Infectious Diseases Research, Department of Microbiology, Faculty of Sciences, Universidad Peruana Cayetano Heredia, Lima, Peru, Asociación Benefica Prisma, Lima, Peru, and Department of International Health, Johns Hopkins Bloomberg School of Public Health, 615 N Wolfe Street, Room W5515, Baltimore, MD 21205. Carlton A. Evans, Department of International Health, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Room W5515, Baltimore, MD 21205, E-mail: caevans@jhsph.edu.

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