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用于定量检测胃泌素-17的增强型发光邻近均相分析方法的开发。

Development of amplified luminescent proximity homogeneous assay for quantitation of gastrin-17.

作者信息

Xiang Zhongyi, Zhou Yunhai, Chen Xindong, Qin Yuan, Zhou Xiumei, Zhao Xueqin, Wang Yigang, Huang Biao, Zhang Yi

机构信息

College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.

Department of General Surgery, The Affiliated Wuxi No.2 People's Hospital of Nanjing Medical University, Wuxi, China.

出版信息

Anal Biochem. 2023 Feb 1;662:115016. doi: 10.1016/j.ab.2022.115016. Epub 2022 Dec 9.

Abstract

A highly sensitive and convenient amplified luminescent proximity homogeneous assay (AlphaLISA) method with high throughput and automation potential was developed for quantitation of serum Gastrin-17 (G-17) levels, which can facilitate the early diagnosis of atrophic gastritis in people at high risk of gastric cancer using a non-invasive approach. In this study, donor and acceptor beads with modified carboxyl groups on the surface were directly coupled to anti-G-17 antibodies through activation was proposed for application in the development of the new AlphaLISA, which can effectively simplify the steps and shorten the reaction time to achieve faster detection. Therefore, the G-17-AlphaLISA only needs to react for 15 min to obtain good analysis results. The proposed method has a wider detection range than commercial enzyme-linked immunosorbent assay (ELISA) kits (0.12-112.8 pmol/L > 0.5-40 pmol/L). In addition, results of G-17-AlphaLISA and ELISA had good correlation and agreement (ρ = 0.936). Importantly, the developed method may be more suitable for the large-scale screening of people at high risk for gastric cancer than traditional ELISA and provides a novel solution for other biomarkers that require accurate, highly sensitive, and high throughput detection.

摘要

开发了一种具有高通量和自动化潜力的高灵敏度、便捷的放大发光邻近均相分析(AlphaLISA)方法,用于定量血清胃泌素-17(G-17)水平,该方法可通过非侵入性方法促进胃癌高危人群萎缩性胃炎的早期诊断。在本研究中,提出将表面带有修饰羧基的供体珠和受体珠通过活化直接偶联至抗G-17抗体,用于新型AlphaLISA的开发,这可有效简化步骤并缩短反应时间以实现更快检测。因此,G-17-AlphaLISA仅需反应15分钟即可获得良好的分析结果。所提出的方法比商业酶联免疫吸附测定(ELISA)试剂盒具有更宽的检测范围(0.12 - 112.8 pmol/L > 0.5 - 40 pmol/L)。此外,G-17-AlphaLISA和ELISA的结果具有良好的相关性和一致性(ρ = 0.936)。重要的是,所开发的方法可能比传统ELISA更适合胃癌高危人群的大规模筛查,并为其他需要准确、高灵敏度和高通量检测的生物标志物提供了一种新的解决方案。

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