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应用原位 PCR、免疫过氧化物酶和 HE 染色技术研究传染性法氏囊病病毒在鸡体内的组织嗜性和感染发病机制。

Infectious bursal disease virus tissue tropism and pathogenesis of the infection in chickens by application of in situ PCR, immunoperoxase and HE staining.

机构信息

Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.

出版信息

Microb Pathog. 2019 Apr;129:195-205. doi: 10.1016/j.micpath.2019.01.049. Epub 2019 Feb 6.

Abstract

Infectious bursal disease is one of an OIE list of notifiable diseases. Chicken is the only host that manifests clinical signs and its pathogenicity is correlated with the distribution of antigens in organs. This study was conducted to determine disease pathogenesis and virus tissue tropism by in situ PCR, immunoperoxidase staining (IPS), and HE staining. Twenty four chickens were infected with very virulent Infectious Bursal Disease Virus (vvIBDV). Fifteen chickens were kept as a control group. Infected chickens were sacrificed at hrs 2, 4, 6, 12, days 1, 2, 4, and 6 post-inoculation (pi). While, control chickens were euthanized on days 0, 1, 2, 4, and 6 pi. Different tissues were collected, fixed in 10% buffered formalin, and processed. At hr 2 pi, virus was detected in intestinal, junction of the proventriculus and gizzard, cecal tonsil, liver, kidney, and bursa of Fabricius. At hr 4 pi, virus reached spleen, and at hr 6 pi, it entered thymus. At hr 12 pi, virus concentration increased in positive tissues. The latest invaded tissue was muscle on day 1 pi. Secondary viraemia occurred during 12-24 h pi. In situ PCR was the most sensitive technique to highlight obscure points of infection in this study.

摘要

传染性法氏囊病是世界动物卫生组织(OIE)规定报告的疾病之一。鸡是唯一表现临床症状的宿主,其致病性与器官中抗原的分布相关。本研究通过原位 PCR、免疫过氧化物酶染色(IPS)和 HE 染色来确定疾病发病机制和病毒组织嗜性。24 只鸡感染了超强毒传染性法氏囊病病毒(vvIBDV)。15 只鸡作为对照组饲养。感染鸡在接种后 2、4、6、12 小时、第 1、2、4 和 6 天被处死。而对照组鸡在接种后第 0、1、2、4 和 6 天被安乐死。收集不同的组织,固定在 10%缓冲福尔马林溶液中,并进行处理。在接种后 2 小时,病毒在肠道、前胃和肌胃交界处、盲肠扁桃体、肝脏、肾脏和法氏囊检测到。在接种后 4 小时,病毒进入脾脏,在接种后 6 小时,病毒进入胸腺。在接种后 12 小时,阳性组织中的病毒浓度增加。最早入侵的组织是第 1 天的肌肉。二次病毒血症发生在接种后 12-24 小时。原位 PCR 是本研究中突出感染隐匿点最敏感的技术。

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