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Parallel detection of multiple zoonotic parasites using a real-time fluorogenic loop-mediated isothermal amplification-based quadruple-sample microfluidic chip.
- Source :
-
Frontiers in microbiology [Front Microbiol] 2023 Sep 26; Vol. 14, pp. 1238376. Date of Electronic Publication: 2023 Sep 26 (Print Publication: 2023). - Publication Year :
- 2023
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Abstract
- Zoonotic parasites pose significant health risks globally. In the present study, we combined a microfluidic chip with loop-mediated isothermal amplification (on-chip LAMP) to detect five zoonotic parasites: Toxoplasma gondii , Cryptosporidium parvum , Cryptosporidium hominis , Clonorchis sinensis , and Taenia solium . This method enabled the simultaneous parallel analysis of five genetic markers from a maximum of four samples per chip. The on-chip LAMP assay was conducted in a highly automated format via the addition (by pipetting) of each sample in a single operation. The reaction was performed in volumes as low as 5 μL at a temperature of 65°C for 60 min, achieving limits of detection ranging from 10 <superscript>-2</superscript> to 10 <superscript>-3</superscript> pg./μL of recombinant plasmid DNA. All the time-to-positive values were less than 40 min, and almost all the coefficients of variation were less than 10%, even when using limit of detection concentrations for multiple pathogens, indicating robust reproducibility among replicates. The clinical sensitivity and specificity for detecting 135 field samples were 98.08 and 97.59%, respectively, compared with traditional biological methods, indicating good applicability in the detection of field samples. This on-chip LAMP assay allows for low reagent consumption, ease of operation, and multiple analyses of samples and genetic targets, and is applicable for on-site detection and the routine monitoring of multiple zoonotic parasites.<br />Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2023 Chen, Lou, Duan, Zhou, Xu, Chen, Chen, Xu, Du and Chen.)
Details
- Language :
- English
- ISSN :
- 1664-302X
- Volume :
- 14
- Database :
- MEDLINE
- Journal :
- Frontiers in microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 37822745
- Full Text :
- https://doi.org/10.3389/fmicb.2023.1238376