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Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
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Parallel detection of multiple zoonotic parasites using a real-time fluorogenic loop-mediated isothermal
Yu-Xin Chen1,2, Yi-Rong Lou1,2, Li-Jun Duan1,2,3
1State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo, China.
Frontiers in Microbiology
|October 12, 2023
Summary
A novel microfluidic chip with loop-mediated isothermal amplification (LAMP) enables rapid, automated detection of five key zoonotic parasites. This on-chip LAMP assay offers high sensitivity and specificity for parasite surveillance.
Area of Science:
- Parasitology
- Molecular Diagnostics
- Microfluidics
Background:
- Zoonotic parasites present a substantial global health challenge.
- Accurate and rapid detection methods are crucial for disease control and prevention.
Purpose of the Study:
- To develop and validate a microfluidic chip-based loop-mediated isothermal amplification (on-chip LAMP) assay.
- To enable simultaneous detection of five major zoonotic parasites: *Toxoplasma gondii*, *Cryptosporidium parvum*, *Cryptosporidium hominis*, *Clonorchis sinensis*, and *Taenia solium*.
Main Methods:
- Integration of microfluidic chip technology with LAMP for parallel analysis of multiple samples and targets.
- Automated sample processing with low reaction volumes (5 μL) and a single incubation temperature (65°C).
- Assessment of limit of detection, time-to-positive, reproducibility, clinical sensitivity, and specificity using recombinant plasmid DNA and field samples.
Main Results:
- Achieved limits of detection from 10⁻² to 10⁻³ pg/μL of recombinant plasmid DNA.
- Demonstrated robust reproducibility with coefficients of variation <10% and time-to-positive values <40 min.
- Reported high clinical sensitivity (98.08%) and specificity (97.59%) for detecting zoonotic parasites in 135 field samples.
Conclusions:
- The developed on-chip LAMP assay is a highly sensitive, specific, and reproducible method for detecting multiple zoonotic parasites.
- The assay's automation, low reagent consumption, and ease of operation make it suitable for on-site detection and routine monitoring.
- This technology offers a significant advancement in the surveillance and control of zoonotic parasitic infections.
