Development and assessment of molecular diagnostic tests for 15 enteropathogens causing childhood diarrhoea: a multicentre study.

Autor:	Liu J; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Kabir F; Department of Pediatrics and Child Health, Aga Khan University, Karachi, Pakistan., Manneh J; Medical Research Council Unit, Banjul, The Gambia., Lertsethtakarn P; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Begum S; International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka, Bangladesh., Gratz J; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA; Kilimanjaro Clinical Research Institute, Moshi, Tanzania., Becker SM; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Operario DJ; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Taniuchi M; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Janaki L; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Platts-Mills JA; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA., Haverstick DM; Department of Pathology, University of Virginia, Charlottesville, VA, USA., Kabir M; International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka, Bangladesh., Sobuz SU; International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka, Bangladesh., Nakjarung K; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Sakpaisal P; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Silapong S; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Bodhidatta L; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Qureshi S; Department of Pediatrics and Child Health, Aga Khan University, Karachi, Pakistan., Kalam A; Department of Pediatrics and Child Health, Aga Khan University, Karachi, Pakistan., Saidi Q; Kilimanjaro Clinical Research Institute, Moshi, Tanzania., Swai N; Kilimanjaro Clinical Research Institute, Moshi, Tanzania., Mujaga B; Haydom Lutheran Hospital, Haydom, Tanzania., Maro A; Kilimanjaro Clinical Research Institute, Moshi, Tanzania., Kwambana B; Medical Research Council Unit, Banjul, The Gambia., Dione M; Medical Research Council Unit, Banjul, The Gambia., Antonio M; Medical Research Council Unit, Banjul, The Gambia., Kibiki G; Kilimanjaro Clinical Research Institute, Moshi, Tanzania., Mason CJ; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand., Haque R; International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka, Bangladesh., Iqbal N; Department of Pediatrics and Child Health, Aga Khan University, Karachi, Pakistan., Zaidi AKM; Department of Pediatrics and Child Health, Aga Khan University, Karachi, Pakistan., Houpt ER; Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, VA, USA. Electronic address: erh6k@virginia.edu.
Jazyk:	angličtina
Zdroj:	The Lancet. Infectious diseases [Lancet Infect Dis] 2014 Aug; Vol. 14 (8), pp. 716-724. Date of Electronic Publication: 2014 Jul 09.
DOI:	10.1016/S1473-3099(14)70808-4
Abstrakt:	Background: Childhood diarrhoea can be caused by many pathogens that are difficult to assay in the laboratory. Molecular diagnostic techniques provide a uniform method to detect and quantify candidate enteropathogens. We aimed to develop and assess molecular tests for identification of enteropathogens and their association with disease. Methods: We developed and assessed molecular diagnostic tests for 15 enteropathogens across three platforms-PCR-Luminex, multiplex real-time PCR, and TaqMan array card-at five laboratories worldwide. We judged the analytical and clinical performance of these molecular techniques against comparator methods (bacterial culture, ELISA, and PCR) using 867 diarrhoeal and 619 non-diarrhoeal stool specimens. We also measured molecular quantities of pathogens to predict the association with diarrhoea, by univariate logistic regression analysis. Findings: The molecular tests showed very good analytical and clinical performance at all five laboratories. Comparator methods had limited sensitivity compared with the molecular techniques (20-85% depending on the target) but good specificity (median 97·3%, IQR 96·5-98·9; mean 95·2%, SD 9·1). Positive samples by comparator methods usually had higher molecular quantities of pathogens than did negative samples, across almost all platforms and for most pathogens (p<0·05). The odds ratio for diarrhoea at a given quantity (measured by quantification cycle, Cq) showed that for most pathogens associated with diarrhoea-including Campylobacter jejuni and Campylobacter coli, Cryptosporidium spp, enteropathogenic Escherichia coli, heat-stable enterotoxigenic E coli, rotavirus, Shigella spp and enteroinvasive E coli, and Vibrio cholerae-the strength of association with diarrhoea increased at higher pathogen loads. For example, Shigella spp at a Cq range of 15-20 had an odds ratio of 8·0 (p<0·0001), but at a Cq range of 25-30 the odds ratio fell to 1·7 (p=0·043). Interpretation: Molecular diagnostic tests can be implemented successfully and with fidelity across laboratories around the world. In the case of diarrhoea, these techniques can detect pathogens with high sensitivity and ascribe diarrhoeal associations based on quantification, including in mixed infections, providing rich and unprecedented measurements of infectious causes. Funding: Bill & Melinda Gates Foundation Next Generation Molecular Diagnostics Project. (Copyright © 2014 Elsevier Ltd. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu