ArticleParasitology research2012
Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini eggs in human fecal samples using a duplex real-time fluorescence resonance energy transfer PCR and melting curve analysis.
Article in Parasitology research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed, 38 citations in OpenAlex.
- Study of the population genetic structure of Opisthorchis-like eggs in northern Thailand using mitochondrial genes.PLoS neglected tropical diseases · 2024Article
- Diagnosis of Human Trematode Infections.Advances in experimental medicine and biology · 2024Review
- Comparing the performance of urine and copro-antigen detection in evaluating Opisthorchis viverrini infection in communities with different transmission levels in Northeast Thailand.PLoS neglected tropical diseases · 2019Article
- Rapid label-free analysis of Opisthorchis viverrini eggs in fecal specimens using confocal Raman spectroscopy.PloS one · 2019Article
- Performance evaluation of existing immunoassays for Clonorchis sinensis infection in China.Parasites & vectors · 2018Article
- LAMPhimerus: A novel LAMP assay for detecting Amphimerus sp. DNA in human stool samples.PLoS neglected tropical diseases · 2017Article
- Development of a PCR Assay for Diagnosing Trematode (Opisthorchis and Haplorchis) Infections in Human Stools.The American journal of tropical medicine and hygiene · 2017Article
- First report and molecular identification of Opisthorchis viverrini infection in human communities from Lower Myanmar.PloS one · 2017Article
- Current status and perspectives of Clonorchis sinensis and clonorchiasis: epidemiology, pathogenesis, omics, prevention and control.Infectious diseases of poverty · 2016Review
- Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections.Clinical microbiology reviews · 2014Review
- Development of a PCR assay and pyrosequencing for identification of important human fish-borne trematodes and its potential use for detection in fecal specimens.Parasites & vectors · 2014Article
- Molecular differentiation of Opisthorchis viverrini and Clonorchis sinensis eggs by multiplex real-time PCR with high resolution melting analysis.The Korean journal of parasitology · 2013Article
- Development of a polymerase chain reaction applicable to rapid and sensitive detection of Clonorchis sinensis eggs in human stool samples.Pathogens and global health · 2013Article
- Diagnosis of early infection and post chemotherapeutic treatment by copro-DNA detection in experimental opisthorchiasis.Parasitology research · 2013Article
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We developed a single step duplex real-time fluorescence resonance energy transfer (FRET) PCR merged with melting curve analysis for the fast detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini eggs in human fecal samples. Two species of mitochondrial NADH dehydrogenase subunit 2 (nad2) DNA elements, the 165-bp nad2 product of C. sinensis and the 209-bp nad2 product of O. viverrini, were amplified by species-specific primers, and the fluorescence melting curve analyses were generated from hybrid of amplicons and two pairs of species-specific fluorophore-labeled probes. By their different fluorescence channels and melting temperatures, both C. sinensis and O. viverrini eggs in infected human fecal samples were detected and differentiated with high (100%) sensitivity and specificity. Detection limit was as little as a single C. sinensis egg and two O. viverrini eggs in 100 mg of fecal sample. The assay could distinguish the DNA of both parasites from the DNA of negative fecal samples and fecal samples with other parasitosis, as well as from the well-defined genomic DNA of human leukocytes and other parasites. It can reduce labor time of microscopic examination and is not prone to carry over contamination of agarose electrophoresis. Our duplex real-time FRET PCR method would be useful to determine the accurate range of endemic areas and/or to discover the co-endemic areas of two liver flukes, C. sinensis and O. viverrini, in Asia. This method also would be helpful for the differential diagnosis of the suspected cases of liver fluke infections among travelers who had visited the endemic countries of those parasites.
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