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.


Journal

Veterinary parasitology
ISSN: 1873-2550
Titre abrégé: Vet Parasitol
Pays: Netherlands
ID NLM: 7602745

Informations de publication

Date de publication:
Apr 2021
Historique:
received: 14 01 2021
revised: 15 02 2021
accepted: 20 02 2021
pubmed: 11 3 2021
medline: 14 8 2021
entrez: 10 3 2021
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 33689962
pii: S0304-4017(21)00054-6
doi: 10.1016/j.vetpar.2021.109394
pii:
doi:

Substances chimiques

Epitopes, B-Lymphocyte 0
Recombinant Proteins 0
Serine Endopeptidases EC 3.4.21.-
hypodermin EC 3.4.21.-
hypodermin C EC 3.4.21.-

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

109394

Informations de copyright

Copyright © 2021 Elsevier B.V. All rights reserved.

Auteurs

Mubeccel Atelge (M)

Department of Parasitology, Faculty of Veterinary Medicine, Kastamonu University, 37150, Kastamonu, Turkey; Vectors and Vector-Borne Diseases Implementation and Research Center, Erciyes University, 38039, Kayseri, Turkey. Electronic address: matelge@kastamonu.edu.tr.

Abdullah Inci (A)

Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, 38039, Kayseri, Turkey; Vectors and Vector-Borne Diseases Implementation and Research Center, Erciyes University, 38039, Kayseri, Turkey.

Alparslan Yildirim (A)

Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, 38039, Kayseri, Turkey; Vectors and Vector-Borne Diseases Implementation and Research Center, Erciyes University, 38039, Kayseri, Turkey.

Ibrahim Sozdutmaz (I)

Vectors and Vector-Borne Diseases Implementation and Research Center, Erciyes University, 38039, Kayseri, Turkey; Department of Virology, Faculty of Veterinary Medicine, Erciyes University, 38039, Kayseri, Turkey.

Peter H Adler (PH)

Department of Plant and Environmental Sciences, Clemson University, Clemson, SC, 29634, USA.

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Classifications MeSH