In:
Frontiers in Microbiology, Frontiers Media SA, Vol. 14 ( 2023-9-26)
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 −2 to 10 −3 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.
Type of Medium:
Online Resource
ISSN:
1664-302X
DOI:
10.3389/fmicb.2023.1238376
DOI:
10.3389/fmicb.2023.1238376.s001
DOI:
10.3389/fmicb.2023.1238376.s002
DOI:
10.3389/fmicb.2023.1238376.s003
Language:
Unknown
Publisher:
Frontiers Media SA
Publication Date:
2023
detail.hit.zdb_id:
2587354-4
