ArticleParasites & vectors2023
A rapid and visual detection assay for Clonorchis sinensis based on recombinase polymerase amplification and lateral flow dipstick.
Article in Parasites & vectors, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 13 citations in OpenAlex.
- A novel rapid detection approach based on CRISPR-Cas13a for Dermatophagoides pteronyssinus and Dermatophagoides farinae (Acariformes: Pyroglyphidae).Journal of insect science (Online) · 2026Article
- High sensitivity detection ofFood science and biotechnology · 2025Article
- Current knowledge on the prevalence and detection techniques ofFrontiers in veterinary science · 2025Review
- Rapid visual detection of Giardia duodenalis in faecal samples using an RPA-CRISPR/Cas12a system.Parasitology research · 2024Article
- A point-of-care testing assay for clonorchiasis using a EuNPs-CsTR1 fluorescent probe-based immunoassay.PLoS neglected tropical diseases · 2024Article
- Application of recombinase polymerase amplification with lateral flow assay to pathogen point-of-care diagnosis.Frontiers in cellular and infection microbiology · 2024Review
- Diagnosis of Human Trematode Infections.Advances in experimental medicine and biology · 2024Review
- Development of an LFD-RPA Assay for Rapid Detection ofCurrent issues in molecular biology · 2023Article
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Authors and funding
11 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundFish-borne zoonotic clonorchiasis, caused by Clonorchis sinensis, is an emerging public health problem in several countries with more than 15 million people infected globally. However, a lack of accurate point-of-care (POC) diagnostic tests in resource-limited areas is still a critical barrier to effective treatment and control of clonorchiasis. The development of the recombinase polymerase amplification(RPA) assay, a POC diagnostic test based on the amplification of pathogen DNA, has provided a new, simple and inexpensive tool for disease detection with high sensitivity and specificity.
methodsA novel RPA method was developed based on specific primers and probes, and combined with the dipstick, to allow for the rapid and intuitive detection of C. sinensis through the amplification of the mitochondrial cytochrome c oxidase subunit 1 (COX1) gene. The lower limit of detection for the combined RPA/lateral flow dipstick (RPA-LFD) assay was evaluated using dilutions of the target DNA sequence. Cross-reactivity was evaluated using genomic DNA from 10 additional control parasites. Forty human clinical stool samples were tested to verify its performance.
resultsThe evaluated primers designed from the C. sinensis COX1 region can be used to detect adult worms, metacercariae, and eggs at 39 °C within 20 min, and the results can be visually observed using the LFD. The detection limit of pathogen genomic DNA was as low as 10 fg, and the number of metacercaria(e) in fish and egg(s) in faeces were both as low as one. This improved the sensitivity of low-infection detection tremendously. The test is species-specific, and no other related control parasites were detected. In human stool samples with eggs per gram (EPG) > 50, the RPA-LFD assay was performed consistent with conventional Kato-Katz (KK) and PCR methods.
conclusionThe established RPA-LFD assay provides a powerful tool for the diagnosis and epidemiological survey of C. sinensis from human and animal samples, and has important implications for the effective control of clonorchiasis.
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