Samir Das, Kandhan Srinivas, Arockiasamy Arun Prince Milton, Sabia Khan, Lavinia Wahlang, Hosterson Kylla, Gundallahalli Bayyappa Manjunatha Reddy, Sharanagouda S Patil, Erica Lawai Lyngdoh, Pebam Chandrima Devi, Sandeep Ghatak, Kekungu-U Puro, Arnab Sen
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引用次数: 0
Abstract
Lumpy Skin Disease (LSD), a viral disease affecting cattle and related species, entered India in 2019, with first outbreak reported in Odisha, and has since caused significant economic losses to the Indian livestock sector. Following its entry, the disease spread rapidly to various states, including the northeastern region which is known for its porous borders. A total of 56 clinical samples (whole blood, skin scrapings, nasal swab, skin lesions and serum) were collected from various outbreaks of LSD in Meghalaya. World Organisation for Animal Health (WOAH)-recommended probe-based real-time polymerase chain reaction (PCR) targeting LSDV126 region detected Lumpy skin disease virus (LSDV) in 42 out of 56 clinical samples. Additionally, 182 contemporary serum samples were screened using commercial ELISA kit which indicated an apparent seroprevalence of 11.54% (95% confidence interval [CI] 7.7-17%) and a true prevalence of 12.3% (95% CI 7.86-18.53%). Three representative samples were subjected to partial sequencing-based phylogenetic analysis targeting the LSDVp32 and LSDV117 regions which confirmed the diagnosis of LSD and also revealed circulation of the Kenyan field strain-associated LSDV. Furthermore, a novel isothermal approach exploiting Saltatory Rolling Circle Amplification (SRCA) mechanism was also explored with WOAH-recommended conventional PCR primers which yielded results comparable to real-time PCR-based diagnostic methods with a minimum detection limit of 10 copies/µL of the standard plasmid. The availability of this rapid and reliable diagnostic assay could be harnessed for early diagnosis of LSD, especially in resource-limited and field laboratory settings and ultimately aid in the timely implementation of control strategies.
肿块性皮肤病(LSD)是一种影响牛及相关物种的病毒性疾病,于2019年进入印度,据报道首次爆发于奥里萨邦,此后给印度畜牧业造成了重大经济损失。该疾病进入后迅速蔓延到各州,包括以边界漏洞百出著称的东北部地区。在梅加拉亚邦的各种LSD暴发中,共收集了56份临床样本(全血、皮肤刮痕、鼻拭子、皮肤病变和血清)。世界动物卫生组织(WOAH)推荐的基于探针的实时聚合酶链反应(PCR)靶向LSDV126区域,在56份临床样本中检测出42份肿块皮肤病病毒(LSDV)。此外,使用商用ELISA试剂盒筛选182份当代血清样本,结果显示表观血清阳性率为11.54%(95%置信区间[CI] 7.7-17%),真实患病率为12.3% (95% CI 7.86-18.53%)。对3份代表性样本进行了以LSDVp32和LSDV117区域为目标的部分测序系统发育分析,证实了LSD的诊断,并揭示了肯尼亚野毒株相关LSDV的循环。此外,研究人员还利用waah推荐的传统PCR引物探索了一种新的等温方法,利用振荡滚动圈扩增(SRCA)机制,其结果与基于实时PCR的诊断方法相当,最低检测限为10拷贝/µL标准质粒。这种快速可靠的诊断方法可用于LSD的早期诊断,特别是在资源有限和现场实验室环境中,并最终有助于及时实施控制策略。
期刊介绍:
AMB Express is a high quality journal that brings together research in the area of Applied and Industrial Microbiology with a particular interest in ''White Biotechnology'' and ''Red Biotechnology''. The emphasis is on processes employing microorganisms, eukaryotic cell cultures or enzymes for the biosynthesis, transformation and degradation of compounds. This includes fine and bulk chemicals, polymeric compounds and enzymes or other proteins. Downstream processes are also considered. Integrated processes combining biochemical and chemical processes are also published.