Development, Optimization and Evaluation of a Sensitive Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detection of Chicken-Based IgY Polyclonal Antibodies against Toxins of D. polylepis Venom.

IF 3 Q3 IMMUNOLOGY
Antibodies Pub Date : 2024-06-21 DOI:10.3390/antib13030050
Stephen Wilson Kpordze, Gideon Mutie Kikuvi, James Hungo Kimotho, Victor Atunga Mobegi
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引用次数: 0

Abstract

Life-threatening medical issues can result from snakebite, and hence this is a public health concern. In many tropical and subtropical nations such as Kenya, where a wide variety of poisonous snakes are prevalent, diagnosis of snakebite in health facilities is imperative. Different antivenoms are needed to treat the venom of different snake species. Nonetheless, it might be difficult for medical professionals to identify the exact snake species that envenomated a patient due to the similarities of several snake envenomations' clinical symptoms. Therefore, the necessity for an assay or technique for identifying venomous species is critical. The current study sought to develop a sensitive ELISA prototype for the detection of D. polylepis venom in Kenya using generated chicken-based IgY polyclonal antibodies. Serum samples containing specific chicken-based IgY antibodies previously raised against D. polylepis venom toxins were used in the assay development. ELISA parameters were optimized, and the developed assay was assessed for applicability. The limit of detection (LoD) of the ELISA for neurotoxic venoms was determined to be 0.01 µg/mL. Successful discrimination between neurotoxic and cytotoxic venoms was achieved by the ensuing inhibition ELISA assay. The developed assay showed the capability of identifying venoms in blood samples (from spiked and venom-challenged blood samples) of BALB/c mice, providing compelling evidence of the strategy's usefulness. This assay could help physicians diagnose and manage victims of snakebites through the evaluation of clinical samples.

开发、优化和评估灵敏的酶联免疫吸附试验(ELISA)原型,用于检测鸡对多刺蛇毒素的 IgY 多克隆抗体。
被蛇咬伤可能导致危及生命的医疗问题,因此这是一个公共卫生问题。在肯尼亚等许多热带和亚热带国家,毒蛇种类繁多,因此必须在医疗机构对蛇咬伤进行诊断。治疗不同种类蛇的毒液需要不同的抗蛇毒血清。然而,由于几种毒蛇咬伤的临床症状相似,医务人员可能很难确定咬伤病人的确切蛇种。因此,必须要有识别毒蛇种类的检测方法或技术。目前的研究试图利用生成的鸡源 IgY 多克隆抗体开发一种灵敏的 ELISA 原型,用于检测肯尼亚的多刺蛇(D. polylepis)毒液。检测开发中使用的血清样本中含有先前针对多刺蛇毒液毒素培养出的特异性鸡基 IgY 抗体。对 ELISA 参数进行了优化,并对所开发的检测方法的适用性进行了评估。ELISA 对神经毒性毒液的检测限 (LoD) 被确定为 0.01 µg/mL。随后的抑制酶联免疫吸附分析法成功区分了神经毒性毒液和细胞毒性毒液。所开发的检测方法显示,它能够识别 BALB/c 小鼠血液样本(来自添加毒液和毒液挑战血液样本)中的毒液,为该策略的实用性提供了令人信服的证据。这种检测方法可以帮助医生通过评估临床样本来诊断和处理蛇咬伤受害者。
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来源期刊
Antibodies
Antibodies IMMUNOLOGY-
CiteScore
7.10
自引率
6.40%
发文量
68
审稿时长
11 weeks
期刊介绍: Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.
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