Molecular identification of<i>Fasciola</i>spp. (Digenea: Platyhelminthes) in cattle from Vietnam

Nguyen, Sam Thi; Amer, Said; Ichikawa, Madoka; Itagaki, Tadashi; Fukuda, Yasuhiro; Nakai, Yutaka

doi:10.1051/parasite/2012191085

Cited by 22 publications

(21 citation statements)

References 27 publications

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“…Neither ITS1 nor ITS2 contained the ambiguous sites, which are typical of hybrid Fasciola specimens. 36 Morphological and morphometric parameters, such as body length to width ratio, and distance between the ventral sucker and the posterior end of the body, also support the identification of our specimen as F. gigantica (S2).…”

Section: Resultssupporting

confidence: 75%

Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay Kit and In-House Fasciola gigantica Cysteine Proteinases-Based Enzyme-Linked Immunosorbent Assays for Diagnosis of Human Fascioliasis

Tran

Intuyod

et al. 2019

The American Journal of Tropical Medicine and Hygiene

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Fascioliasis, caused by Fasciola hepatica and Fasciola gigantica infection, is a major food-borne trematodiasis in many places of the world, with the central region of Vietnam being reported as a highly endemic area. Stool examination for Fasciola eggs is not a sensitive method, and immunodiagnostic methods are preferable. We investigated various enzyme-linked immunosorbent assays (ELISAs) to evaluate their efficacy for fascioliasis diagnosis. Test sera used are primarily screened using an ELISA kit produced in Vietnam (VN kit; Viet Sinh Chemical Producing & Trading Co. Ltd., Ho Chi Minh City, Vietnam): Seropositive individuals having symptoms compatible with fascioliasis were regarded as clinically diagnosed fascioliasis cases. A commercial Fasciola IgG ELISA kit from Diagnostic Automation/Cortez Diagnostics, Inc. (USA kit; Woodland Hills, CA), which has been commonly used in Vietnam, was assessed and compared with in-house ELISA systems, including a cystatincapture (CC) ELISA using crude worm extract (CWE) and an indirect ELISA using a synthetic peptide Ac-TPTCHWECQVGYNKTYDEE-NHMe designed from the F. gigantica cathepsin B (FgCB5) molecule. The USA kit was suitable for routine diagnosis after recalibration of the manufacturer's suggested cutoff point. Cystatin-capture ELISA with CWE provided good sensitivity and specificity with perfect agreement to the results of the USA kit. In dotblot ELISA, recombinant FgCB5 reacted more strongly with human antisera than did other F. gigantica antigens tested. Enzyme-linked immunosorbent assay using the synthetic peptide fragment of the FgCB5 exhibited nearly 80% sensitivity and specificity, but the test results showed low agreement with CC-ELISA or the USA kit. In conclusion, the commercially available Fasciola IgG ELISA kit from the United States and the in-house CC ELISA using CWE are suitable for practical diagnosis for fascioliasis.

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Section: Resultssupporting

confidence: 75%

Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay Kit and In-House Fasciola gigantica Cysteine Proteinases-Based Enzyme-Linked Immunosorbent Assays for Diagnosis of Human Fascioliasis

Tran

Intuyod

et al. 2019

The American Journal of Tropical Medicine and Hygiene

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“…Most human cases have been reported from Binh Dinh, Phu Yen, and Khanh Hoa Provinces in Central Vietnam, which are associated with local endemic animal fascioliasis (33,34,35,47). Recent studies of cattle in Binh Dinh indicated an overall prevalence of 45.3% for Fasciola eggs using a sedimentation method (33,34), 54.9% using a coprological approach, and 72.2% by serological analysis (35). Fascioliasis is clearly hyperendemic in cattle in Vietnam, with attendant risks for the human population.…”

Section: Discussionmentioning

confidence: 99%

Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)

Nguyen

et al. 2012

J Clin Microbiol

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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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“…It is estimated that 2.4 million people in more than 60 countries are infected, and the number of people at risk is more than 180 million throughout the world (1). F. hepatica infects more than 300 million cattle and 250 million sheep worldwide (2). F. hepatica and F. gigantica causes considerable economic losses to worldwide farming estimated as more than US$ 3.2 billion annually (3,4).…”

Section: Introductionmentioning

confidence: 99%

Morphometric and Molecular Study of Fasciola Isolates from Ruminants in Iran

Akhlaghi¹,

Mohammadi²,

Ziaali³

et al. 2018

TurkiyeParazitolDerg

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Objective: The purpose of the present study was morphometric and molecular characterization of Fasciola isolates from ruminants in Iran. Methods: Flukes were collected from the livers of 54 naturally infected sheep and cattle. The proportion of body length to width (L/W) of each fresh fluke was measured using a digital caliper. We employed receiver operating characteristic (ROC) curve analysis to explore the reliability of L/W for differentiating the two species. Polymerase chain reaction (PCR)-sequencing was performed on ribosomal Internal Transcribed Spacers (ITS) and mitochondrial cytochrome c oxidase subunit 1 (cox1) genes. The sequences were then analyzed and phylogenetic relationships were investigated. Result: Forty-eight out of 54 isolates (88.9%) were identified as F. hepatica and four isolates (7.4%) as F. gigantica. All the sheep isolates were F. hepatica, while 4 out of 10 cattle were infected with F. gigantica. The morphometric study revealed an L/W ratio of 1.2 to 6.5 in Fasciola isolates with significantly higher L/W ratio in F. gigantica (p<0.00). According to the ROC curve analysis, the L/W value of 3.55 was regarded as the critical value to discriminate between the two species. Conclusions: Findings of the present study indicate the presence of both Fasciola species in southeastern Iran. The phylogenetic analysis revealed two different clades representing F. hepatica and F. gigantica. The two isolates in this study were described as Fasciola sp. The mitochondrial DNA of these isolates were similar to F. hepatica, while their ITS fragments were identical to F. gigantica.

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Molecular identification ofFasciolaspp. (Digenea: Platyhelminthes) in cattle from Vietnam

Cited by 22 publications

References 27 publications

Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay Kit and In-House Fasciola gigantica Cysteine Proteinases-Based Enzyme-Linked Immunosorbent Assays for Diagnosis of Human Fascioliasis

Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay Kit and In-House Fasciola gigantica Cysteine Proteinases-Based Enzyme-Linked Immunosorbent Assays for Diagnosis of Human Fascioliasis

Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)

Morphometric and Molecular Study of Fasciola Isolates from Ruminants in Iran

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