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First molecular characterization of hypodermin genes of Hypoderma bovis and serodiagnosis of bovine hypodermosis with recombinant hypodermin C antigen and a synthetic peptide containing its linear B-cell epitope      
Yazarlar
Dr. Öğr. Üyesi Mübeccel ATELGE Dr. Öğr. Üyesi Mübeccel ATELGE
Kastamonu Üniversitesi, Türkiye
Abdullah İnci
Erciyes Üniversitesi, Türkiye
Alparslan Yıldırım
Erciyes Üniversitesi, Türkiye
İbrahim Sözdutmaz
Türkiye
Peter H. Adler
Özet
Hypodermins A (HA), B (HB), and C (HC) of warble flies are modulatory antigens involved in host inflammation and immune responses during migration of the warble fly larvae through host connective tissues. In the current study, molecular characteristics of the genes encoding HA, HB, and HC were revealed from cDNA constructs of third-instar larvae of Hypoderma bovis. The open reading frame (ORF) of each hypodermin gene was amplified with modified gene-specific primers, and the resulting PCR products were cloned into pGEM-T Easy Vector to produce recombinant plasmids (rHA, rHB, and rHC). The ORF sequences of rHA, rHB, and rHC genes are 705 bp, 771 bp, and 783 bp long and encode proteins of 234, 256, and 263 amino acids with predicted sizes of 25.74 kDa, 27.79 kDa, and 28.51 kDa, respectively. The rHC gene was subcloned into the pET 100/D-TOPO Expression Vector, and the recombinant HC was purified using affinity chromatography. Western blotting indicated that rHC was recognized by the sera of cattle naturally infested with H. bovis. The rHC and a synthetic peptide (sHC) containing its linear B cell-specific epitope were evaluated as serological markers in indirect ELISA (iELISA) for the diagnosis of bovine hypodermosis. Both sHC and rHC iELISAs had sensitivity values equal to or higher than 90 % and specificity values of 100 %. A total of 200 serum samples from cattle in the Central Anatolia Region of Turkey were also analyzed by rHC and sHC-iELISAs to reveal the seroprevalence of bovine hypodermosis. The results of both iELISAs were consistent with one another and revealed a hypodermosis prevalence of 62 %. Our study provides the first data on molecular characterization of hypodermin genes of H. bovis and indicates the efficacy of recombinant antigen and peptide-based iELISA for serodiagnosis of bovine hypodermosis.
Anahtar Kelimeler
Hypoderma bovis | Hypodermin genes | Molecular characterization | Recombinant antigen | Serodiagnosis | Synthetic peptide
Makale Türü Özgün Makale
Makale Alt Türü SSCI, AHCI, SCI, SCI-Exp dergilerinde yayımlanan tam makale
Dergi Adı VETERINARY PARASITOLOGY
Dergi ISSN 0304-4017
Dergi Tarandığı Indeksler SCI-Expanded
Dergi Grubu Q1
Makale Dili Türkçe
Basım Tarihi 04-2021
Cilt No 292
Sayı 292
Doi Numarası 10.1016/j.vetpar.2021.109394
Makale Linki http://dx.doi.org/10.1016/j.vetpar.2021.109394