Isolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR

dc.creatorMagalhães, Kelly Grace
dc.creatorJannotti-Passos, Liana K
dc.creatorCaldeira, Roberta Lima
dc.creatorBerne, Maria Elisabeth Aires
dc.creatorMuller, Gertrude
dc.creatorCarvalho, Omar dos Santos
dc.creatorLenzi, Henrique Leonel
dc.date2012-07-17T18:32:06Z
dc.date2012-07-17T18:32:06Z
dc.date2008
dc.date.accessioned2023-09-26T20:07:46Z
dc.date.available2023-09-26T20:07:46Z
dc.identifierMAGALHÃES, K. G. et al. Isolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR. Veterinary Parasitology, v. 152, n.3-4, p. 333-8, 2008.
dc.identifier0304-4017
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/4173
dc.identifier10.1016/j.vetpar.2007.12.019
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8849204
dc.descriptionDetection of Fasciola hepatica infection in Lymnaea viatrix through analysis of histological cuts is based upon morphological characters of the parasite during the intra-mollusk phase of parasitism. At this stage, trematode forms are very similar and, thus, very difficult to differentiate. Specific detection may also be impaired by the presence of other helminthes in the mollusk. Histological samples are usually fixed in formalin, embedded in paraffin, sectioned and HE stained. In the current study, a method for the extraction of DNA from formalin-fixed, paraffin-embedded tissues was standardized by means of deparaffinizing with xylol and digesting with proteinase K. Extracted DNA was amplified in a multiplex-PCR, by using simultaneous primers in a single reaction under high stringency conditions. Results showed specific amplification of DNA from the trematode and the snails. The technique was sensitive enough to detect F. hepatica infections in L. viatrix, in histological sections in which the presence of larval stages could not be observed through brightfield microscopy. The profiles generated were: stair bands referring to F. hepatica DNAmt amplification; a band of 1200 bp referring to L. viatrix ITS and another of 1300 bp referring to F. hepatica ITS and other trematodes. Multiplex-PCR has shown to be a fast, safe, highly sensitive and specific method, which is able to amplify DNA from fixed tissues, despite a low DNA quantity and its degradation caused by fixation processes. Such methodology may be useful in studies on fascioliasis epidemiology, enabling the use of material from histological collections.
dc.formatapplication/pdf
dc.languageeng
dc.publisherElsevier
dc.rightsrestricted access
dc.subjectFasciola hepatica
dc.subjectDetection
dc.subjectMultiplex-PCR
dc.subjectDNA isolation
dc.subjectFormalin-fixed
dc.subjectParaffin-embedded
dc.subjectPrimers do DNA
dc.subjectDNA de Helmintos
dc.subjectFasciola hepatica
dc.subjectVetores de Doenças
dc.subjectReservatórios de Doenças
dc.titleIsolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR
dc.typeArticle


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