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理性截短适体用于跨物种检测眼镜蛇咬伤。

Rational truncation of aptamer for cross-species application to detect krait envenomation.

机构信息

Department of Biotechnology, All India Institute of Medical Sciences (AIIMS), New Delhi, 110029, India.

Faculty of Pharmacy, Uttarakhand Technical University (UTU), Dehradun, 248007, Uttarakhand, India.

出版信息

Sci Rep. 2018 Dec 12;8(1):17795. doi: 10.1038/s41598-018-35985-1.

Abstract

In majority of snakebite cases, the snake responsible for the bite remains unidentified. The traditional snakebite diagnostics method relies upon clinical symptoms and blood coagulation assays that do not provide accurate diagnosis which is important for epidemiological as well as diagnostics point of view. On the other hand, high batch-to-batch variations in antibody performance limit its application for diagnostic assays. In recent years, nucleic acid aptamers have emerged as a strong chemical rival of antibodies due to several obvious advantages, including but not limited to in vitro generation, synthetic nature, ease of functionalization, high stability and adaptability to various diagnostic formats. In the current study, we have rationally truncated an aptamer developed for α-Toxin of Bungarus multicinctus and demonstrated its utility for the detection of venom of Bungarus caeruleus. The truncated aptamer α-Tox-T2 (26mer) is found to have greater affinity than its 40-mer parent counterpart α-Tox-FL. The truncated aptamers are characterized and compared with parent aptamer for their binding, selectivity, affinity, alteration in secondary structure and limit of detection. Altogether, our findings establish the cross-species application of a DNA aptamer generated for α-Toxin of Bungarus multicinctus (a snake found in Taiwan and China) for the reliable detection of venom of Bungarus caeruleus (a snake found in the Indian subcontinent).

摘要

在大多数蛇伤病例中,负责咬伤的蛇仍然无法确定。传统的蛇伤诊断方法依赖于临床症状和凝血检测,但这些方法并不能提供准确的诊断,这在流行病学和诊断学方面都很重要。另一方面,抗体性能在批间存在很大差异,限制了其在诊断检测中的应用。近年来,核酸适体作为抗体的有力化学替代品出现,具有几个明显的优势,包括但不限于体外产生、合成性质、易于功能化、高稳定性和对各种诊断形式的适应性。在本研究中,我们合理地截短了针对 Bungarus multicinctus 的α-毒素的适体,并证明了它在检测 Bungarus caeruleus 毒液中的应用。截短的适体α-Tox-T2(26 个碱基)比其 40 个碱基的母体对应物α-Tox-FL 具有更高的亲和力。对截短的适体及其母体适体进行了结合、选择性、亲和力、二级结构改变和检测限的比较。总之,我们的研究结果确立了为台湾和中国发现的 Bungarus multicinctus 的α-毒素生成的 DNA 适体在可靠检测印度次大陆发现的 Bungarus caeruleus 毒液方面的跨物种应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b16/6290766/2b66096a947e/41598_2018_35985_Fig1_HTML.jpg

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