A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica

Marcilla, Antonio; Bargues, M. D.; Mas‐Coma, Santiago

doi:10.1006/mcpr.2002.0429

Cited by 129 publications

(107 citation statements)

References 22 publications

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“…Different molecular techniques have been used for the characterization of liver fukes. Concerning the mitochondrial DNA, part of the mt-DNA of F. hepatica was sequenced and showed length heterogeneity, suggesting length differences among individual mitochondrial genomes (Marcilla et al, 2002). In the present study, the mt-DNA CO1 gene digestion patterns differed markedly between F. hepatica and F. gigantica.…”

Section: Resultsmentioning

confidence: 44%

“…The differentiation becomes very difficult when dealing with small specimens. At the extremes of this morphological range, some resemble F. hepatica, whereas others resemble F. gigantica, with intermediate forms also occurring and involving phenomena such as abnormal gametogenesis, diploidy, triploidy and mixoploidy, parthenogenesis, and hybridization events between different genotypes (Mas-Coma & Bargues, 1997;Marcilla et al, 2002). The two species can also be discriminated by DNA sequences of nuclear ribosomal internal transcribed spacer 1 (ITS-1), ITS-2, and 28S rRNA genes (Adlard et al, 1993;Marcilla et al, 2002;Itagaki & Tsutsumi, 1998;Itagaki et al, 2005) and of mitochondrial NDI and CO1 genes (Itagaki et al, 2005 …”

Section: Introductionmentioning

confidence: 99%

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Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica

Şimşek

Ütük²,

Balkaya

2011

Helminthologia

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The most common and widespread liver flukes of the genus Fasciola are Fasciola hepatica and F. gigantica. Adults of both species occur in many domestic ruminants and in humans and can cause serious disease. The differential diagnosis of these flukes infection is very important because of their different transmission and epidemiological characteristics. A simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the common restriction enzymes AluI and RsaI, is described to distinguish between both fasciolid species. After the digestion of the mitochondrial cytochrome c oxidase 1 (CO1) PCR product with the restriction enzyme AluI, the RFLP profile obtained from F. hepatica revealed two fragments, whereas F. gigantica was not cut. The RsaI digestion generated two fragments from F. gigantica, whereas it did not cut the PCR product from F. hepatica. Results were confirmed with CO1 sequence analysis of both F. hepatica and F. gigantica. The present study suggests that the PCRRFLP method described here can be used for the proper identification of Fasciola species.

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Section: Resultsmentioning

confidence: 44%

Section: Introductionmentioning

confidence: 99%

Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica

Şimşek

Ütük²,

Balkaya

2011

Helminthologia

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“…With respect to F. hepatica researches PCR-RFLP has been often utilized in different cases and for various genes. Marcilla et al (2002) (Huang et al, 2003), ITS 1 (internal transcribed spacer 1) of rDNA or cox1 (cytochrome c oxidase I) mitochondrial gene (Hashimoto et al, 1997) as genetic markers. Through PCR-RFLP analysis Walker et al (2007) have defined for F. hepatica three mitochondrial DNA (mtDNA) regions with highest polymorphism frequency.…”

Section: Pcr-rflp (Restriction Fragment Length Polymorphism)mentioning

confidence: 99%

“…SNP assay of that fragment sequence revealed intraspecific genetic variability. Sequence analysis again of 28S rDNA region in combination with PCR-RFLP assay also allowed differentiation between F. hepatica and F. gigantica (Marcilla et al, 2002). Although intraspecific polymorphisms have not been proved and only a few iterspecific variation have been found.…”

Section: Ribosomal Dna Regions Used For Snp Assaymentioning

confidence: 99%

Issues Associated with Genetic Diversity Studies of the Liver Fluke, Fasciola Heptica (Platyhelminthes, Digenea, Fasciolidae)

Teofanova¹,

Hristov²,

Yoveva³

et al. 2012

Genetic Diversity in Microorganisms

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“…A more limited study, developed in Turkey, showed the diversity of F. hepatica strains by sequencing the mitochondrial gene nad1 and the ribosomal part ITS1 (Dosay-Akbulut et al, 2005). Other previous studies (Marcilla et al, 2002;Ramadan & Saber, 2004) were developed to distinguish F. hepatica from F. gigantica, particularly in countries where both species are sympatric. Intra-specific and inter-specific comparisons were made between parasites from cattle and/or sheep.…”

Section: Diversity Of Parasites In the Paragonimus Genusmentioning

confidence: 99%

Biodiversity of flukes

Dreyfuss

Rondelaud

2008

Parasite

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Summary :As many others parasites, speciation of flukes depends on the genetic characteristics and on ploidia. Ploidia of flukes can be different in a same species. In Asia, diploid, triploid and hybrid (2n/3n) populations are encountered. The comparison of morphological parameters between diploid and triploid flukes showed that they were morphologically different. Nevertheless, a genetic relationship between parthenogenetic organisms would exist regardless of their ploidia. In the Fasciola genus, the main consequence of the high level of diversity is the frequent probability of development of resistance to anthelmintics and fast adaptation to climatic changes. In the Paragonimus genus, diversity can enhance different forms of pathogenicity, can also be related to the species of intermediate hosts, and to the definitive host. The strain of flukes plays a part in the visceral localization of P. westermani adults. DIFFICULTY OF SPECIATION IN MEDICAL PARASITOLOGYPLOIDIA P arasitology is a biological field in which taxonomy often still uses morphological and Linnean criteria. The limits of the Linnean descriptive methods were reached many decades ago: the dilemma Entamoeba histolytica/E. dispar is a demonstrative example. Similarly, the infra-specific relationships between biogeography of parasites and their biological characteristics (rhythm, pathology, reservoirs) were used to organize the taxonomy of genetically different parasitic populations (Wuchereria bancrofti, for instance). Genetic diversity cannot yet be evaluated by easy and low cost methods. The results of the biological diagnosis of parasitic diseases tend to be less precise, and decrease at the genus level (E. histolytica/E. dispar), or even the family (Opisthorchiidae, Ancylostomatidae, Paragonimidae). Protozoa are parasites for which genetic diversity can be evaluated quite easily. Helminths, particularly Digenea, cannot be analyzed easily, because many species have a variable ploidia, which modifies their morphology and DNA size. A comparative morphometric study, by light microscopy, of eggs and adults from three populations with different sizes of Fasciola gigantica, and the morphology of adults, under scanning electron microscopy, showed variations in the sizes of eggs and adults, whereas the surface was similar using electronic morphology (Srimuzipo et al., 2000). The observations on mitotic metaphase chromosomes revealed a diploid type (2n = 20) in all three size-races. The speciation of Digenea can no longer be evaluated by simple morpholo-

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A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica

Cited by 129 publications

References 22 publications

Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica

Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica

Issues Associated with Genetic Diversity Studies of the Liver Fluke, Fasciola Heptica (Platyhelminthes, Digenea, Fasciolidae)

Biodiversity of flukes

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