Cloning and expression of Fasciola hepatica enolase gene and efficacy of recombinant protein in the serodiagnosis of sheep fasciolosis

Basit Öğe Kaydı
dc.contributor.authorCelik, Figen
dc.contributor.authorSimsek, Sami
dc.contributor.authorSelcuk, Muhammed Ahmed
dc.contributor.authorKesik, Harun Kaya
dc.contributor.authorKilinc, Seyma Gunyakti
dc.contributor.authorCelik, Burcak Aslan
dc.date.accessioned2024-12-24T19:27:48Z
dc.date.available2024-12-24T19:27:48Z
dc.date.issued2023
dc.departmentSiirt Üniversitesi
dc.description.abstractFasciolosis caused by Fasciola hepatica is a disease of zoonotic importance that is common worldwide and can cause serious problems in farm animals, some wild animals and humans. The development of diagnostic kits for the correct diagnosis of fasciolosis in sheep is important in terms of preventing yield losses. With this study, it is aimed to clone and express the enolase gene to be isolated from adult F. hepatica and to determine the effectiveness of the recombinant antigen in the serodiagnosis of sheep fasciolosis. For this aim, primers were designed to amplify the enolase gene from the F. hepatica enolase sequence, mRNA was isolated from F. hepatica adult fluke obtained from an infected sheep followed by cDNA was obtained. Enolase gene was amplified by PCR and the product was cloned and then expressed. The efficiency of the purified recombinant protein was displayed by Western blot (WB) and ELISA using positive and negative sheep sera. As a result, the sensitivity and specificity rates of the recombinant FhENO antigen were 85% and %82.8 by WB while the rates were 90% and 97.14% by ELISA, respectively. At the same time, in sheep blood sera samples collected from the Elazig and Siirt provinces of Turkey, 100 (50%) of 200 sera were found to be positive by WB and 46 (23%) were found to be positive by ELISA. The most important problem in ELISA was the high cross-reaction rate of the recombinant antigen used, as in WB. In order to prevent the cross-reactions, it will be useful to compare the genes encoding the enolase protein of parasites from the closely related parasite family, and select the regions where there are no common epitopes, and clone them and test the purified protein.
dc.description.sponsorshipScientific and Technological Research Council of Turkey (TUBITAK) [121O789]
dc.description.sponsorshipThis study was financially supported by a grant (121O789) from the Scientific and Technological Research Council of Turkey (TUBITAK). This study was part of the first author's PhD thesis, named Cloning and Expression of Fasciola hepatica Enolase Gene and Determination of Efficacy of Recombinant Protein in the Diagnosis of Sheep Fasciolosis presented at Firat University
dc.identifier.doi10.1016/j.vetpar.2023.109961
dc.identifier.issn0304-4017
dc.identifier.issn1873-2550
dc.identifier.pmid37290212
dc.identifier.scopus2-s2.0-85161054042
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.1016/j.vetpar.2023.109961
dc.identifier.urihttps://hdl.handle.net/20.500.12604/6801
dc.identifier.volume320
dc.identifier.wosWOS:001019110900001
dc.identifier.wosqualityQ2
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherElsevier
dc.relation.ispartofVeterinary Parasitology
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_20241222
dc.subjectFasciola hepatica
dc.subjectSheep
dc.subjectEnolase
dc.subjectCloning
dc.subjectWestern Blot
dc.subjectELISA
dc.titleCloning and expression of Fasciola hepatica enolase gene and efficacy of recombinant protein in the serodiagnosis of sheep fasciolosis
dc.typeArticle

Dosyalar

Koleksiyon

WOS İndeksli Yayınlar Koleksiyonu
PubMed İndeksli Yayın Koleksiyonu
Scopus İndeksli Yayınlar Koleksiyonu