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猪德尔塔冠状病毒重组膜蛋白候选抗原基因的合成及其用于检测血清阳性猪的研究

A Candidate Antigen of the Recombinant Membrane Protein Derived from the Porcine Deltacoronavirus Synthetic Gene to Detect Seropositive Pigs.

机构信息

Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Km 15.5 Carretera México-Toluca, Palo Alto, Cuajimalpa, Ciudad de México 05110, Mexico.

Posgrado en Ciencias de la Producción y de la Salud Animal, Facultad de Estudios Superiores Cuautitlán, Estado de México, Universidad Nacional Autónoma de México, Ciudad de México 04510, Mexico.

出版信息

Viruses. 2023 Apr 25;15(5):1049. doi: 10.3390/v15051049.

Abstract

Porcine deltacoronavirus (PDCoV) is an emergent swine coronavirus which infects cells from the small intestine and induces watery diarrhea, vomiting and dehydration, causing mortality in piglets (>40%). The aim of this study was to evaluate the antigenicity and immunogenicity of the recombinant membrane protein (M) of PDCoV (M-PDCoV), which was developed from a synthetic gene obtained after an in silico analysis with a group of 138 GenBank sequences. A 3D model and phylogenetic analysis confirmed the highly conserved M protein structure. Therefore, the synthetic gene was successfully cloned in a pETSUMO vector and transformed in BL21 (DE3). The M-PDCoV was confirmed by SDS-PAGE and Western blot with ~37.7 kDa. The M-PDCoV immunogenicity was evaluated in immunized (BLAB/c) mice and iELISA. The data showed increased antibodies from 7 days until 28 days ( < 0.001). The M-PDCoV antigenicity was analyzed using pig sera samples from three states located in "El Bajío" Mexico and positive sera were determined. Our results show that PDCoV has continued circulating on pig farms in Mexico since the first report in 2019; therefore, the impact of PDCoV on the swine industry could be higher than reported in other studies.

摘要

猪德尔塔冠状病毒(PDCoV)是一种新兴的猪冠状病毒,感染小肠细胞并引起水样腹泻、呕吐和脱水,导致仔猪死亡率(>40%)。本研究旨在评估 PDCoV 重组膜蛋白(M-PDCoV)的抗原性和免疫原性,该蛋白源自通过对来自 138 个 GenBank 序列的组进行计算机分析获得的合成基因。3D 模型和系统发育分析证实了 M 蛋白结构高度保守。因此,成功地将合成基因克隆到 pETSUMO 载体中,并转化到 BL21(DE3)中。通过 SDS-PAGE 和 Western blot 验证了 M-PDCoV,其分子量约为 37.7 kDa。通过免疫接种(BLAB/c)小鼠和 iELISA 评估了 M-PDCoV 的免疫原性。数据显示,从第 7 天到第 28 天抗体增加(<0.001)。使用来自墨西哥“El Bajío”三个州的猪血清样本分析了 M-PDCoV 的抗原性,并确定了阳性血清。我们的结果表明,自 2019 年首次报告以来,PDCoV 一直在墨西哥的养猪场中持续传播;因此,PDCoV 对养猪业的影响可能高于其他研究报告的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cca1/10222885/23f8dde7bf2a/viruses-15-01049-g001.jpg

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