Development of an ELISA for the detection of fowl adenovirus serotype −4 utilizing fiber protein

IF 1.5 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Biologicals Pub Date : 2024-02-01 Epub Date: 2024-02-23 DOI:10.1016/j.biologicals.2024.101752
Faiza Ather , Muhammad Ashir Zia , Mudasser Habib , Muhammad Salah-ud-din Shah
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Abstract

Hydropericardium syndrome (HPS), caused by the Fowl adenovirus 4 (FAdV-4) has led to significant financial losses for the poultry industry globally, including Pakistan over the past few years. Conventional serological methods are time consuming, laborious and less sensitive therefore, a rapid and sensitive ELISA kit is required for the reliable detection of FAdV-4 infection. In the current research, fiber proteins (1 &2) of FAdV-4 were successfully expressed in Escherichia coli and purified using metal affinity chromatography. Using these proteins as antigens, an indirect ELISA for detecting FAdV-4 infection was developed. The developed ELISA showed superior performances upon comparison with Serum neutralization test (SNT). This ELISA also showed reliable detection of FAdV specific antibodies in experimentally infected and vaccinated chickens. This assay produced good correlation on the samples collected from the field with SNT and found essential for large scale serology of the FAdV. No cross reactivity was observed in the ELISA following the testing of the serum samples of different other avian pathogens which showed that this ELISA is specific in detecting the FAdV infection. In conclusion, the developed Fiber protein ELISA is highly sensitive and specific in the detecting the FAdV infection and can be utilized for large scale sero-epidemiology of the disease.

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开发一种利用纤维蛋白检测鸡腺病毒血清型-4 的 ELISA 方法
在过去几年里,由鸡腺病毒 4(FAdV-4)引起的心包积水综合症(HPS)给包括巴基斯坦在内的全球家禽业造成了巨大的经济损失。传统的血清学方法费时、费力且灵敏度较低,因此需要一种快速灵敏的 ELISA 试剂盒来可靠地检测 FAdV-4 感染。目前的研究成功地在大肠杆菌中表达了 FAdV-4 的纤维蛋白(1 &2 ),并使用金属亲和层析法进行了纯化。以这些蛋白为抗原,开发了一种检测 FAdV-4 感染的间接 ELISA。与血清中和试验(SNT)相比,所开发的酶联免疫吸附试验(ELISA)表现出更优越的性能。该酶联免疫吸附试验还能可靠地检测实验感染鸡和疫苗接种鸡的 FAdV 特异性抗体。该检测法与 SNT 检测法在现场采集的样本中产生了良好的相关性,被认为是大规模 FAdV 血清学检测的必要方法。在检测其他不同禽类病原体的血清样本后,ELISA 没有发现交叉反应,这表明该 ELISA 在检测 FAdV 感染方面具有特异性。总之,所开发的纤维蛋白 ELISA 在检测 FAdV 感染方面具有高度灵敏性和特异性,可用于该疾病的大规模血清流行病学研究。
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来源期刊
Biologicals
Biologicals 生物-生化研究方法
CiteScore
3.70
自引率
0.00%
发文量
39
审稿时长
48 days
期刊介绍: Biologicals provides a modern and multidisciplinary international forum for news, debate, and original research on all aspects of biologicals used in human and veterinary medicine. The journal publishes original papers, reviews, and letters relevant to the development, production, quality control, and standardization of biological derived from both novel and established biotechnologies. Special issues are produced to reflect topics of particular international interest and concern.Three types of papers are welcome: original research reports, short papers, and review articles. The journal will also publish comments and letters to the editor, book reviews, meeting reports and information on regulatory issues.
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