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开发一种作为家禽新城疫快速诊断检测的协同凝集试剂盒。

Development of a coagglutination kit as a rapid test for diagnosing Newcastle disease in poultry.

作者信息

Naf'an Muhammad Kholish, Kurniasih Kurniasih, Untari Tri, Prakoso Yos Adi

机构信息

Student of Master of Sciences Degree, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, Indonesia.

Department of Pathology, Faculty of Veterinary Medicine, University of Gadjah Mada, Yogyakarta, Indonesia.

出版信息

Vet World. 2020 Aug;13(8):1719-1724. doi: 10.14202/vetworld.2020.1719-1724. Epub 2020 Aug 28.

DOI:10.14202/vetworld.2020.1719-1724
PMID:33061250
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7522963/
Abstract

BACKGROUND AND AIM

Newcastle disease (ND) is a viral infection that causes high mortality and economic loss in the poultry industry. The Office International des Epizooties (OIE) recommends several diagnostic methods for the detection of ND, including isolation and molecular tests. However, these detection methods are time-consuming and highly expensive. Therefore, this study was conducted to develop a coagglutination kit as a novel diagnostic tool for ND in the poultry industry.

MATERIALS AND METHODS

Two adult male New Zealand White rabbits weighing 2.5 kg were vaccinated using ND life vaccine intraperitoneally. The vaccination was conducted once a week for 4 weeks with multilevel doses. Rabbits' serum was collected at week 6 and inactivated at 56°C for 30 min. The serum was precipitated using ammonium sulfate and reacted with protein A of Staphylococcus aureus to produce the agglutination kit for detecting ND virus. A total of 25 chickens suspected with ND infection from a local poultry farm in Yogyakarta were used as the test samples. The chickens were necropsied, and the brain, spleen, lung, intestine, and feces were collected. Half of these organs were subjected to tests using the coagglutination kit and reverse transcription-polymerase chain reaction (RT-PCR). The other half was processed for histopathology. Data were analyzed qualitatively.

RESULTS

Of the 25 samples, 13 (52%) were positive for ND infection when tested using both the ND coagglutination kit and RT-PCR. The positive samples also exhibited several histopathological changes, including perivascular cuffing surrounding the cerebral blood-brain barrier, hemorrhagic pneumonia, splenitis, and necrotic hemorrhage enteritis.

CONCLUSION

This study confirmed that the ND coagglutination kit could be used as a novel diagnostic tool for the detection of ND virus infection in the poultry industry.

摘要

背景与目的

新城疫(ND)是一种病毒感染,可在家禽业中导致高死亡率和经济损失。国际兽疫局(OIE)推荐了几种用于检测新城疫的诊断方法,包括分离和分子检测。然而,这些检测方法耗时且成本高昂。因此,本研究旨在开发一种协同凝集试剂盒,作为家禽业中新城疫的新型诊断工具。

材料与方法

选用两只体重2.5千克的成年雄性新西兰白兔,通过腹腔注射新城疫活疫苗进行免疫接种。每周接种一次,共接种4周,采用多剂量水平。在第6周采集兔子血清,并在56℃灭活30分钟。血清用硫酸铵沉淀,与金黄色葡萄球菌蛋白A反应,制备用于检测新城疫病毒的凝集试剂盒。从日惹当地一家家禽养殖场选取25只疑似感染新城疫的鸡作为测试样本。对这些鸡进行剖检,收集脑、脾、肺、肠和粪便。其中一半器官使用协同凝集试剂盒和逆转录聚合酶链反应(RT-PCR)进行检测。另一半进行组织病理学处理。对数据进行定性分析。

结果

在25个样本中,使用新城疫协同凝集试剂盒和RT-PCR检测时,有13个(52%)样本新城疫感染呈阳性。阳性样本还表现出一些组织病理学变化,包括围绕脑血脑屏障的血管周围套袖现象、出血性肺炎、脾炎和坏死性出血性肠炎。

结论

本研究证实,新城疫协同凝集试剂盒可作为家禽业中检测新城疫病毒感染的新型诊断工具。

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