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A multiplex PCR-based sequencing method for the diagnosis and differentiation of echinococcosis using plasma samples,
Yanping Zhao1, Yeqin Wang2, Ming Zhi3
1BGI Research, Shenzhen 518083, China; BGI Research, Chongqing 401329, China.
Cell Reports Methods
|May 8, 2026
Summary
A new multiplex PCR test detects Echinococcus cell-free DNA (cfDNA) in plasma, offering a low-cost, non-invasive method for diagnosing echinococcosis and differentiating species. This approach shows promise for identifying the parasitic infection accurately.
Area of Science:
- Medical Diagnostics
- Parasitology
- Molecular Biology
Background:
- Echinococcosis is a significant public health concern, typically diagnosed using imaging techniques.
- Current diagnostic methods can be invasive or lack specificity.
- Non-invasive detection of Echinococcus cell-free DNA (cfDNA) in plasma presents a promising alternative.
Purpose of the Study:
- To develop and validate a low-cost multiplex PCR panel for the non-invasive detection and species differentiation of Echinococcus from plasma cfDNA.
- To assess the sensitivity and specificity of the developed PCR panel in clinical samples.
- To evaluate the correlation between cfDNA levels and echinococcosis lesion size.
Main Methods:
- Development of a multiplex PCR panel with 230 primer pairs targeting Echinococcus mitochondrial and nuclear DNA.
- Validation using serial dilutions of pathogen DNA and 81 human plasma samples (53 patients, 28 controls).
- Species differentiation through sequencing analysis of PCR amplicons.
Main Results:
- The assay detected Echinococcus genomic DNA at inputs as low as 1 fg/reaction.
- Clinical sample testing yielded a sensitivity of 68.42% and specificity of 92.86%.
- 80% of clinically confirmed echinococcosis cases were correctly identified; cfDNA levels correlated with lesion size.
Conclusions:
- Targeted cfDNA sequencing offers a potent tool for non-invasive echinococcosis diagnosis and species identification.
- The developed multiplex PCR panel is a cost-effective method with potential for use in clinical settings.
- This approach can aid in diagnosing challenging cases and monitoring disease burden.