从孟加拉国牛群中分离、适应和鉴定结节性皮肤病病毒
Isolation, adaptation, and characterization of lumpy skin disease virus from cattle in Bangladesh.
作者信息
Pervin Sumaiya, Ahamed Md Mostakin, Chouhan Chandra Shaker, Jahan Md Salim, Ahmed Rony, Nazmul K H M, Nazir Hussain, Siddique Mahbubul Pratik, Rahman Md Tanvir, Kafi Md Abdul, Rahman Md Bahanur
机构信息
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh.
Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh, Bangladesh.
出版信息
J Adv Vet Anim Res. 2023 Sep 30;10(3):563-569. doi: 10.5455/javar.2023.j710. eCollection 2023 Sep.
OBJECTIVE
The research aimed to isolate, adapt to cell culture, and characterize the lumpy skin disease virus (LSDV) from clinically infected cattle in Bangladesh.
MATERIALS AND METHODS
From September 2019 to June 2020, 37 skin nodules and skin swabs were aseptically collected from afflicted cattle in the outbreak regions of Jhenaidah and Kishoreganj in Bangladesh. The LSDV was isolated from embryonated specific pathogen-free (SPF) chicken eggs along the chorioallantoic membrane (CAM) route and the Vero cell line after several blind passages. The viral attachment protein was targeted for molecular detection using polymerase chain reactions (PCR). For phylogenetic analysis, PCR-positive products were partially sequenced.
RESULTS
The virus was evident in the cell line, showed cytopathic effects after the 13 blind passage, and on the CAM of SPF chicken eggs, exhibited thickening of the CAM with pock-like lesions. A total of 12 samples (32.43%) tested positive for LSDV by PCR. Phylogenetic analysis of the present isolates (accession numbers MN792649 and MN792650) revealed 100% similarity with strains from India (MN295064), Kenya (AF325528, MN072619, KX683219), Greece (KY829023), Serbia (KY702007), and Kazakhstan (MN642592); moreover, 99.43% to 100% similarity to the sheep pox virus.
CONCLUSION
Partially sequenced LSDV was developed as a vaccine seed and was first isolated in Bangladesh and characterized at the molecular level.
目的
本研究旨在从孟加拉国临床感染牛中分离出结节性皮肤病病毒（LSDV），使其适应细胞培养并进行特性鉴定。
材料与方法
2019年9月至2020年6月，从孟加拉国杰纳伊达和基肖尔甘杰疫情爆发地区的患病牛身上无菌采集了37个皮肤结节和皮肤拭子。经过多次盲传后，通过绒毛尿囊膜（CAM）途径从无特定病原体（SPF）鸡胚以及Vero细胞系中分离出LSDV。使用聚合酶链反应（PCR）对病毒附着蛋白进行分子检测。为进行系统发育分析，对PCR阳性产物进行了部分测序。
结果
该病毒在细胞系中明显可见，在第13次盲传后出现细胞病变效应，在SPF鸡胚的CAM上，表现为CAM增厚并伴有痘样病变。共有12个样本（32.43%）通过PCR检测LSDV呈阳性。对当前分离株（登录号MN792649和MN792650）的系统发育分析显示，与来自印度（MN295064）、肯尼亚（AF325528、MN072619、KX683219）、希腊（KY829023）、塞尔维亚（KY702007）和哈萨克斯坦（MN642592）的毒株具有100%的相似性；此外，与绵羊痘病毒的相似性为99.43%至100%。
结论
部分测序的LSDV被开发为疫苗种子，首次在孟加拉国分离并在分子水平上进行了特性鉴定。
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