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.

Novel approaches to the diagnosis of Strongyloides stercoralis infection

“…Also, although not only, for this reason, diagnostic methods that are not dependent on skilled microscopists are needed; therefore, the application of molecular assays is expected to increase in parasitology, which is still lagging behind in this respect, as compared with virology or bacteriology. With the relatively recent advent of microarray technology, and the advances in lab-on-chip systems, potentially allowing the simultaneous and fully automated detection of multiple pathogens [86], this will become even more true. However, in the particular case of S. stercoralis infection, molecular diagnosis has yet to demonstrate an optimal sensitivity, which is particularly required for this parasite, for which even very light infections are relevant and must be detected and treated.…”

Lab-on-Chip-Based Platform for Fast Molecular Diagnosis of Multidrug-Resistant Tuberculosis

“…In addition, the LoC can be adapted for other diagnostic or research needs, thus providing a S531L S315T1 WT 1 D516V S315T1 T-8C D516V S315T1 T-8C 1 WT WT WT WT WT WT 15 WT S315T1 WT WT S315T1 WT 1 S S WT WT WT 9 pos WT WT WT WT 4 WT WT WT PCNV PCNV PCNV 1 S S ND S315T1 WT MTBND MTBND MTBND 1 S S ND WT WT ND ND ND 1 multipurpose platform suitable for other relevant diseases (e.g., influenza, malaria, tropical diseases) (25,26). We thank Tanja Ubben and Tanja Struwe Sonnenschein for excellent technical assistance and Enrico Tortoli for valuable support.…”

A Novel Pan-FlavivirusDetection and Identification Assay Based on RT-qPCR and Microarray

“…Previously published microarray or diagnostic chip approaches [20] lack the ease of operation of the ArrayStrip platform used here and cover only part of the range of viruses that we can identify [12, 14, 19, 46]. …”