设计一种可用于动物致病性钩端螺旋体感染的重组多表位DNA疫苗

Q4 Veterinary World''s Veterinary Journal Pub Date : 2022-12-25 DOI:10.54203/scil.2022.wvj44
B. Garba, N. I. Dirie
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引用次数: 0

摘要

钩端螺旋体病可导致人类严重疾病和可能的死亡。来自致病菌株的抗原表位已显示出作为候选疫苗的潜力,并在免疫诊断测试的敏感性和特异性中发挥关键作用。进行这种体外分析是为了使用计算机表位预测方法开发原型重组DNA疫苗。表位预测软件程序预测了OmpL1、LipL32、LipL41、Loa22和LigA的最具抗原性的线性B细胞表位。预测了13个表位,通过Gly-Ser连接子连接并合成。通过限制性内切酶消化和凝胶电泳评估浓缩的重组多表位蛋白的纯度。在哺乳动物中国仓鼠卵巢细胞系上的体外表达表明,基于间接免疫荧光抗体测试产生的强细胞质荧光。细胞质的绿色表示根据间接免疫荧光抗体测试在未转染的对照组中观察到的针对蓝色染色的细胞核的成功转录和翻译的DNA。目前的研究结果表明,疫苗构建体具有很高的抗体结合潜力,可用于诊断应用或作为多价候选疫苗。
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Designing a Recombinant Multi-epitope DNA Vaccine as Candidate for Protection against Pathogenic Leptospira Infection in Animals
Leptospirosis can cause severe disease and probable death in humans. Antigenic epitopes from pathogenic strains of the bacteria have shown potential for serving as vaccine candidates and play a key role in the sensitivity and specificity of immunodiagnostic tests. This in-vitro analysis was undertaken to develop a prototype recombinant DNA vaccine using in-silico epitope prediction method. Epitope prediction software programs predicted the most antigenic linear B-cell epitopes of OmpL1, LipL32, LipL41, Loa22, and LigA. Thirteen epitopes were predicted, connected by the Gly-Ser linker, and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by restriction enzyme digestion and gel electrophoresis. In-vitro expression on mammalian Chinese Hamster Ovary cell line indicated strong cytoplasmic fluorescence produced based on an indirect immunofluorescence antibody test. The green color of the cytoplasm indicates successful transcribed and translated DNA as against the blue-stained nucleus observed in the un-transfected control group based on the indirect immunofluorescence antibody test. The findings of the current study showed high antibody binding potentials of the vaccine constructs, which could be used for diagnostic applications or as polyvalent vaccine candidates.
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来源期刊
World''s Veterinary Journal
World''s Veterinary Journal Veterinary-Veterinary (all)
CiteScore
1.00
自引率
0.00%
发文量
43
期刊介绍: The World''s Veterinary Journal (ISSN 2322-4568) is an international, peer reviewed open access journal aims to publish the high quality material from veterinary scientists'' studies. All accepted articles are published Quarterly in full text on the Internet. WVJ publishes the results of original scientific researches, reviews, case reports and short communications, in all fields of veterinary science. In details, topics are: Behavior Environment and welfare Animal reproduction and production Parasitology Endocrinology Microbiology Immunology Pathology Pharmacology Epidemiology Molecular biology Immunogenetics Surgery Virology Physiology Vaccination Gynecology Exotic animals Animal diseases Radiology Ophthalmology Dermatology Chronic disease Anatomy Non-surgical pathology issues of small to large animals Cardiology and oncology.
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