title: An integrated lab-on-chip for rapid identification and simultaneous differentiation of tropical pathogens.
abstract: Tropical pathogens often cause febrile illnesses in humans and are responsible for considerable morbidity and mortality. The similarities in clinical symptoms provoked by these pathogens make diagnosis difficult. Thus, early, rapid and accurate diagnosis will be crucial in patient management and in the control of these diseases. In this study, a microfluidic lab-on-chip integrating multiplex molecular amplification and DNA microarray hybridization was developed for simultaneous detection and species differentiation of 26 globally important tropical pathogens. The analytical performance of the lab-on-chip for each pathogen ranged from 102 to 103 DNA or RNA copies. Assay performance was further verified with human whole blood spiked with Plasmodium falciparum and Chikungunya virus that yielded a range of detection from 200 to 4×105 parasites, and from 250 to 4×107 PFU respectively. This lab-on-chip was subsequently assessed and evaluated using 170 retrospective patient specimens in Singapore and Thailand. The lab-on-chip had a detection sensitivity of 83.1% and a specificity of 100% for P. falciparum; a sensitivity of 91.3% and a specificity of 99.3% for P. vivax; a positive 90.0% agreement and a specificity of 100% for Chikungunya virus; and a positive 85.0% agreement and a specificity of 100% for Dengue virus serotype 3 with reference methods conducted on the samples. Results suggested the practicality of an amplification microarray-based approach in a field setting for high-throughput detection and identification of tropical pathogens.
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.
authors: Jeslin J L
authors: JJ
authors: Tan
authors: CI Group, Molecular Diagnostic Business Unit, Microfluidics Division, ST Microelectronics, Catania, Italy.
authors: Monica
authors: M
authors: Capozzoli
authors: Veredus Laboratories Pte Ltd, Singapore Science Park, Singapore.
authors: Mitsuharu
authors: M
authors: Sato
authors: Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Mae Sot, Thailand.
authors: Wanitda
authors: W
authors: Watthanaworawit
authors: Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Mae Sot, Thailand.
authors: Clare L
authors: CL
authors: Ling
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.
authors: Marjorie
authors: M
authors: Mauduit
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.
authors: Benoît
authors: B
authors: Malleret
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.
authors: Anne-Charlotte
authors: AC
authors: Grüner
authors: Veredus Laboratories Pte Ltd, Singapore Science Park, Singapore.
authors: Rosemary
authors: R
authors: Tan
authors: Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Mae Sot, Thailand; Centre for Tropical Medicine, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors: François H
authors: FH
authors: Nosten
authors: Université Pierre et Marie Curie (Paris VI), Centre Hospitalo-Universitaire Pitié-Salpêtrière, Paris, France; INSERM UMR S 945, Paris, France.
authors: Georges
authors: G
authors: Snounou
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore.
authors: Laurent
authors: L
authors: Rénia
authors: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore; Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors: Lisa F P
authors: LF
authors: Ng
mesh_terms: D003141:Communicable Diseases / Q000175:diagnosis*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D012846:Singapore; D013785:Thailand; D014330:Tropical Medicine / Q000295:instrumentation / Q000379:methods*