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埃及禽呼肠孤病毒(ARV)的分子与病理学研究以及与疫苗株相比的田间毒株遗传变异性评估。

Molecular and pathological investigation of avian reovirus (ARV) in Egypt with the assessment of the genetic variability of field strains compared to vaccine strains.

作者信息

Mosad Samah M, Elmahallawy Ehab Kotb, Alghamdi Abeer M, El-Khayat Fares, El-Khadragy Manal F, Ali Lobna A, Abdo Walied

机构信息

Department of Virology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.

Department of Zoonoses, Faculty of Veterinary Medicine, Sohag University, Sohag, Egypt.

出版信息

Front Microbiol. 2023 Apr 17;14:1156251. doi: 10.3389/fmicb.2023.1156251. eCollection 2023.

Abstract

Avian orthoreovirus (ARV) is among the important viruses that cause drastic economic losses in the Egyptian poultry industry. Despite regular vaccination of breeder birds, a high prevalence of ARV infection in broilers has been noted in recent years. However, no reports have revealed the genetic and antigenic characteristics of Egyptian field ARV and vaccines used against it. Thus, this study was conducted to detect the molecular nature of emerging ARV strains in broiler chickens suffering from arthritis and tenosynovitis in comparison to vaccine strains. Synovial fluid samples ( = 400) were collected from 40 commercial broiler flocks in the Gharbia governorate, Egypt, and then pooled to obtain 40 samples, which were then used to screen ARV using reverse transcriptase polymerase chain reaction (RT-PCR) with the partial amplification of ARV sigma C gene. The obtained RT-PCR products were then sequenced, and their nucleotide and deduced amino acid sequences were analyzed together with other ARV field and vaccine strains from GenBank. RT-PCR successfully amplified the predicted 940 bp PCR products from all tested samples. The phylogenetic tree revealed that the analyzed ARV strains were clustered into six genotypic clusters and six protein clusters, with high antigenic diversity between the genotypic clusters. Surprisingly, our isolates were genetically different from vaccine strains, which aligned in genotypic cluster I/protein cluster I, while our strains were aligned in genotypic cluster V/protein cluster V. More importantly, our strains were highly divergent from vaccine strains used in Egypt, with 55.09-56.23% diversity. Sequence analysis using BioEdit software revealed high genetic and protein diversity between our isolates and vaccine strains (397/797 nucleotide substitutions and 148-149/265 amino acid substitutions). This high genetic diversity explains the vaccination failure and recurrent circulation of ARV in Egypt. The present data highlight the need to formulate a new effective vaccine from locally isolated ARV strains after a thorough screening of the molecular nature of circulating ARV in Egypt.

摘要

禽正呼肠孤病毒(ARV)是导致埃及家禽业遭受巨大经济损失的重要病毒之一。尽管种鸡定期接种疫苗,但近年来肉鸡中ARV感染的患病率仍很高。然而,尚无报告揭示埃及田间ARV的遗传和抗原特性以及针对它所使用的疫苗。因此,本研究旨在检测与疫苗株相比,患有关节炎和腱鞘炎的肉鸡中新兴ARV毒株的分子特性。从埃及加比亚省的40个商业肉鸡群中收集了400份滑液样本,然后将其合并以获得40个样本,随后使用逆转录聚合酶链反应(RT-PCR)对ARV的sigma C基因进行部分扩增来筛选ARV。然后对获得的RT-PCR产物进行测序,并将其核苷酸和推导的氨基酸序列与来自GenBank的其他ARV田间和疫苗株一起进行分析。RT-PCR成功地从所有测试样本中扩增出预测的940 bp PCR产物。系统发育树显示,所分析的ARV毒株聚为六个基因型簇和六个蛋白簇,基因型簇之间具有高度的抗原多样性。令人惊讶的是,我们的分离株在基因上与疫苗株不同,疫苗株在基因型簇I/蛋白簇I中,而我们的毒株在基因型簇V/蛋白簇V中。更重要的是,我们的毒株与埃及使用的疫苗株高度不同,差异为55.09-56.23%。使用BioEdit软件进行的序列分析显示,我们的分离株与疫苗株之间存在高度的遗传和蛋白多样性(397/797个核苷酸替换和148-149/265个氨基酸替换)。这种高度的遗传多样性解释了埃及ARV疫苗接种失败和反复传播的原因。目前的数据突出表明,在对埃及循环ARV的分子特性进行全面筛选后,需要从本地分离的ARV毒株中研制一种新的有效疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/30fb/10150020/5a6f0bd41cf8/fmicb-14-1156251-g0001.jpg

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