ArticleJournal of clinical microbiology2012
Development and evaluation of a single-step duplex PCR for simultaneous detection of Fasciola hepatica and Fasciola gigantica (family Fasciolidae, class Trematoda, phylum Platyhelminthes).
Article in Journal of clinical microbiology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
21 citing papers in PubMed, 55 citations in OpenAlex.
- Advancement in Diagnosis, Treatment, and Vaccines againstPathogens (Basel, Switzerland) · 2024Review
- Comparative evaluation of real-time PCR and ELISA for the detection of human fascioliasis.Scientific reports · 2024Article
- Diagnosis of Human Trematode Infections.Advances in experimental medicine and biology · 2024Review
- Fascioliasis.Advances in experimental medicine and biology · 2024Article
- Low-cost molecular methods to characterise gastrointestinal nematode co-infections of goats in Africa.Parasites & vectors · 2023Article
- Ethanolic extract ofOpen veterinary journal · 2023Article
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- High prevalence of liver fluke infestation,Open veterinary journal · 2023Article
- New perspectives for fascioliasis in Upper Egypt's new endemic region: Sociodemographic characteristics and phylogenetic analysis of Fasciola in humans, animals, and lymnaeid vectors.PLoS neglected tropical diseases · 2022Article
- Evaluation of Semi-Nested PCR Compared with Indirect-ELISA to Diagnose Human Fasciolosis.Iranian journal of public health · 2022Article
- Environmental detection ofPeerJ · 2022Article
- Fasciola hepatica and Fasciola gigantica coexistence in domestic ruminants in Nigeria: application of a PCR-based tool.Tropical animal health and production · 2020Article
- New insight into genetic variation and haplotype diversity of Fasciola hepatica from Algeria.Parasitology research · 2019Article
- High-Resolution Melting Analysis as an Appropriate Method to Differentiate betweenIranian journal of public health · 2019Article
- Fasciola and fasciolosis in ruminants in Europe: Identifying research needs.Transboundary and emerging diseases · 2018Review
- Scrambled eggs: A highly sensitive molecular diagnostic workflow for Fasciola species specific detection from faecal samples.PLoS neglected tropical diseases · 2017Article
- Recombinase Polymerase Amplification Compared to Real-Time Polymerase Chain Reaction Test for the Detection ofThe American journal of tropical medicine and hygiene · 2017Article
- Estimating trematode prevalence in snail hosts using a single-step duplex PCR: how badly does cercarial shedding underestimate infection rates?Parasites & vectors · 2014Article
- Molecular testing for clinical diagnosis and epidemiological investigations of intestinal parasitic infections.Clinical microbiology reviews · 2014Review
- Development of two antibody detection enzyme-linked immunosorbent assays for serodiagnosis of human chronic fascioliasis.Journal of clinical microbiology · 2014Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of Fasciola hepatica and F. gigantica. These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of cox1-trnT-rrnL, two species-specific forward primers were designed, FHF (for F. hepatica) and FGF (for F. gigantica), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of Fasciola DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for F. hepatica and 615 bp for F. gigantica. The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each Fasciola species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32 Fasciola samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of Fasciola species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of Fasciola species in areas of distributional and zonal overlap.
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