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Am. J. Trop. Med. Hyg., 79(6), 2008, pp. 893-898
Copyright © 2008 by The American Society of Tropical Medicine and Hygiene

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Diagnosis of Pediatric Pulmonary Tuberculosis by Stool PCR

Hilary Wolf, Melissa Mendez, Robert H. Gilman, Patricia Sheen, Giselle Soto, Angie K. Velarde, Mirko Zimic, A. Roderick Escombe, Sonia Montenegro, Richard A. Oberhelman, AND Carlton A. Evans*
Asociación Benéfica Prisma, Lima, Perú; Laboratorios de Investigación y Desarrollo, Facultad de Ciencias y Filosofia, Universidad Peruana Cayetano Heredia, Lima, Perú; Johns Hopkins Bloomberg School of Public Health, Baltimore; Faculty of Medicine, Universidad de Concepción, Concepción, Chile; Tulane School of Public Health and Tropical Medicine, Tulane, Louisiana; Wellcome Centre for Clinical Tropical Medicine and Department of Infectious Diseases and Immunity; Imperial College London Hammersmith Hospital Campus, London, United Kingdom

Pediatric pulmonary tuberculosis diagnosis is difficult because young children are unable to expectorate sputum samples. Testing stool for tuberculosis DNA from swallowed sputum may diagnose pulmonary tuberculosis. Hospitalized children with suspected tuberculosis had stool, nasopharyngeal, and gastric aspirates cultured that confirmed pulmonary tuberculosis in 16/236 patients. Twenty-eight stored stools from these 16 children were used to evaluate stool polymerase chain reaction (PCR) for tuberculosis diagnosis compared with 28 stool samples from 23 healthy control children. Two DNA extraction techniques were used: fast-DNA mechanical homogenization and Chelex-resin chemical extraction. DNA was tested for tuberculosis DNA with a hemi-nested IS6110 PCR. PCR after Fast-DNA processing was positive for 6/16 culture-proven tuberculosis patients versus 5/16 after Chelex extraction (sensitivity 38% and 31%, respectively). All controls were negative (specificity 100%). If sensitivity can be increased, stool PCR would be a rapid, non-invasive, and relatively bio-secure initial test for children with suspected pulmonary tuberculosis.

Received November 13, 2007. Accepted for publication July 29, 2008.

Acknowledgments: We thank the numerous professional colleagues whose generous collaboration made this work possible and the patients who kindly agreed to give informed consent to provide gastric aspirates, nasopharyngeal aspirates, and stool samples in the interests of medical progress.

Financial support: This study was funded by NIH Project 1 RO1 AI49139-01A1, The National Institute of Allergy and Infectious Diseases; Wellcome Trust Clinical Fellowships in Tropical Medicine; the Global Research Training grant; and the charity Innovation For Health And Development (IFHAD).

Disclosure: The authors declare that they have no conflict of interests in relation to this work.

* Address correspondence to Carlton A. Evans, Asociación Benefica PRISMA, Av Carlos Gonzales Nro 251, Urb. Maranga, San Miguel, Lima 32, Peru. E-mail: CarltonEvans{at}yahoo.com

Authors’ addresses: Hilary Wolf and Giselle Soto, Asociacion Benefica PRISMA, Av Carlos Gonzales Nro 251, Urbanizacion Maranga, San Miguel, Lima 32, Peru, Tel/Fax: 51-1-464-0221. Robert H. Gilman and Carlton A. Evans, Johns Hopkins Bloomberg School of Public Health, 615 N Wolfe Street, w5515, Baltimore, MD 21205, Tel: 1-410-614-3639 or -3959, Fax: 1-410-614-6060. Melissa Mendez, Patricia Sheen, Angie K. Velarde, and Mirko Zimic, Laboratorio de Investigacion y Desarollo, Departamento de Microbiologia, Universidad Peruana Cayetano Heredia, Av. Honorio Delgado 430, San Martin de Porras, Lima, Peru, Tel: 51-1-483-2942, Fax: 51-1-482-4541. A. Roderick Escombe, Department of Infectious Diseases and Immunity, Imperial College London, Hammersmith Hospital Campus, London W12 0NN, UK, Tel: 44-20-8383-3222, Fax: 44-20-8383-3394. Sonia Montenegro, Faculty of Medicine, University of Concepcion, Concepcion, Chile. Richard A. Oberhelman, Dept. of Tropical Medicine, SL-29, 1440 Canal Street, Suite 2210, New Orleans, LA 70112, Tel: 504-988-2511, Fax: 504-988-7313, E-mail: oberhel{at}tulane.edu.

Reprint requests: Carlton A Evans, Asociación Benefica PRISMA, Av Carlos Gonzales Nro 251, Urb. Maranga, San Miguel, Lima 32, Peru, E-mail: CarltonEvans{at}yahoo.com.




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