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一步双重逆转录聚合酶链反应法诊断犊牛腹泻中的A组牛轮状病毒、牛冠状病毒、K99菌属、菌属:组织病理学和免疫层析法

Diagnosis of bovine group A rotavirus, bovine coronavirus, K99, spp., spp. in calf diarrhea with one-step duplex RT-PCR: histopathological and immunochromotographic methods.

作者信息

Irehan Bunyamin, Ozkaraca Mustafa, Gulactı Irem, Sener Levent, Dursun Muhammed Fatih

机构信息

Department of Parasitology, Faculty of Veterinary Medicine, University of Harran, Şanliurfa, Türkiye.

Department of Pathology, Faculty of Veterinary Medicine, University of Sivas Cumhuriyet, Sivas, Türkiye.

出版信息

Vet Res Forum. 2025;16(4):221-227. doi: 10.30466/vrf.2024.2025299.4208. Epub 2025 Apr 15.

DOI:10.30466/vrf.2024.2025299.4208
PMID:40400737
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12091263/
Abstract

Infections causing diarrhea in cattle breeding pose a substantial threat to calf breeding and health, causing considerable economic losses worldwide, including in Türkiye. The primary causative agents of calf diarrhea include bovine group A rotavirus, bovine coronavirus (BCoV), K99, spp., and spp. The objective of this study was to diagnose the pathogens present in samples taken from calves exhibiting diarrhea symptoms in the Elaziğ province of Türkiye. The study material consisted of 85 fecal samples from calves up to 30 days of age with diarrhea symptoms and 42 small intestine samples from calves died as a result of diarrhea symptoms after necropsy. Following total RNA isolation from the samples, one-step duplex reverse transcription polymerase chain reaction (RT-PCR) analysis was conducted to identify the bovine rotavirus (BRV) and BCoV. Our results indicated that 24 of the samples were positive for BRV, 13 were positive for BCoV, and 25 were positive for BRV and BCoV in the form of mixed infection. The immunochromatographic analysis revealed that three samples were positive for K99 antigens, 12 for spp. antigens, and 28 for spp. antigens. Our results indicate that one-step duplex RT-PCR, immunohistochemistry, and immunochromatography methods can be valuable tests to be used in routine diagnostic laboratories for the detection of pathogens associated with calf diarrhea.

摘要

在养牛业中,引起腹泻的感染对犊牛养殖和健康构成了重大威胁,在包括土耳其在内的全球范围内造成了相当大的经济损失。犊牛腹泻的主要病原体包括牛A组轮状病毒、牛冠状病毒(BCoV)、K99、某些菌属和某些菌属。本研究的目的是诊断从土耳其埃拉泽省出现腹泻症状的犊牛采集的样本中存在的病原体。研究材料包括85份来自30日龄以下有腹泻症状犊牛的粪便样本,以及42份在尸检后因腹泻症状死亡的犊牛的小肠样本。从样本中提取总RNA后,进行一步双重逆转录聚合酶链反应(RT-PCR)分析以鉴定牛轮状病毒(BRV)和BCoV。我们的结果表明,24份样本BRV呈阳性,13份样本BCoV呈阳性,25份样本以混合感染的形式BRV和BCoV均呈阳性。免疫层析分析显示,3份样本K99抗原呈阳性,12份样本某些菌属抗原呈阳性,28份样本某些菌属抗原呈阳性。我们的结果表明,一步双重RT-PCR、免疫组织化学和免疫层析方法可作为常规诊断实验室用于检测与犊牛腹泻相关病原体的有价值检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/d82ff485d2a8/vrf-16-221-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/b779157aabd1/vrf-16-221-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/ec276c67f597/vrf-16-221-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/d82ff485d2a8/vrf-16-221-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/b779157aabd1/vrf-16-221-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/ec276c67f597/vrf-16-221-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7996/12091263/d82ff485d2a8/vrf-16-221-g003.jpg

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