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首例基于重组瘦尾蛹Hypodermin C 的 Przhevalskiana silenus 间接 ELISA 在山羊狂蝇蛆病血清学诊断中的应用。

First report of Przhevalskiana silenus derived recombinant hypodermin C based indirect ELISA for serodiagnosis of goat warble fly myiasis.

机构信息

Division of Veterinary Parasitology, Faculty of Veterinary Sciences and Animal Husbandry, Sher-E-Kashmir University of Agricultural Sciences and Technology of Jammu, RS Pura, Jammu, Union Territory of Jammu and Kashmir, 181102, India.

Division of Veterinary Pathology, Faculty of Veterinary Sciences and Animal Husbandry, Sher-E-Kashmir University of Agricultural Sciences and Technology of Jammu, RS Pura, Jammu, Union Territory of Jammu and Kashmir, 181102, India.

出版信息

Sci Rep. 2022 Aug 4;12(1):13440. doi: 10.1038/s41598-022-17760-5.

Abstract

Goat warble fly infestation (GWFI) is a subcutaneous myiasis caused by larvae of Przhevalskiana silenus, an insect belonging to the order Diptera. The diagnosis of GWFI is challenging in the early larval instars (L1 and L2) as they are occult under the skin and hair coat causing prolonged economic loss in form of meat and hide damage. This necessitates early diagnosis for disease control at herd level and its prophylactic management to prevent economic losses. Hypodermins, a class of serine proteases from Hypoderminae subfamily have been used as serodiagnostic antigens for the past four decades for diagnosis of warble fly myiasis. In this study,the immunodominant antigen Hypodermin C (HyC) from P. silenus has been recombinantly expressed in E. coli and immunogenic characterisation of expressed protein was done. The protein shows hallmark residues in conserved cysteine and catalytic triad typical of serine proteases along with similar profile of immunoreactivity towards Hypoderminae infestation. The present study reports an optimised indirect-ELISA based on recombinant HyC derived from P. silenus for early diagnosis of GWFI. The optimised indirect ELISA provides a sensitive and specific immunodiagnostic for mass surveillance of the GWFI with diagnostic specificity and sensitivity of 96% and 100%, respectively and not showing any cross reactivity against other important parasitic and bacterial diseases of goats. This study presents the first report of indirect ELISA based on recombinant Hypodermin C antigen derived from P. silenus for the serosurveillance of goat warble fly disease.

摘要

羊狂蝇蛆病(GWFI)是由幼虫 Przhevalskiana silenus 引起的皮下蝇蛆病,该幼虫属于双翅目昆虫。GWFI 的早期幼虫期(L1 和 L2)的诊断具有挑战性,因为它们隐藏在皮肤和毛发下,导致肉和皮的损害,从而造成长期的经济损失。这就需要在群体水平上进行早期诊断和预防性管理,以控制疾病并防止经济损失。Hypodermins 是 Hypoderminae 亚科的一类丝氨酸蛋白酶,在过去的四十年中一直被用作狂蝇蛆病的血清学诊断抗原。在这项研究中,来自 P. silenus 的免疫显性抗原 Hypodermin C(HyC)在大肠杆菌中被重组表达,并对表达蛋白进行了免疫原性特征分析。该蛋白显示出保守半胱氨酸和催化三联体的标志性残基,这些残基是丝氨酸蛋白酶的典型特征,同时对 Hypoderminae 感染具有相似的免疫反应特征。本研究报告了一种基于重组 P. silenus HyC 的优化间接 ELISA,用于 GWFI 的早期诊断。优化后的间接 ELISA 提供了一种敏感且特异的免疫诊断方法,可用于 GWFI 的大规模监测,其诊断特异性和敏感性分别为 96%和 100%,并且与其他重要的山羊寄生虫和细菌性疾病没有交叉反应。本研究首次报道了基于重组 Hypodermin C 抗原的间接 ELISA 用于山羊狂蝇病的血清学监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0ad/9352896/ebe767a2f0a4/41598_2022_17760_Fig1_HTML.jpg

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