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乙型肝炎病毒检测的等温核酸扩增方法的研究进展。

Advances in isothermal nucleic acid amplification methods for hepatitis B virus detection.

机构信息

State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, Chengdu, China.

Clinical Laboratory, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.

出版信息

Analyst. 2023 Aug 7;148(16):3708-3718. doi: 10.1039/d3an00700f.

Abstract

Hepatitis B virus (HBV) infection is a major global health problem of widespread concern. Clinically, serological assays are the most widely used diagnostic tests for HBV infection, with the presence of HBsAg in the serum being indicative of acute and chronic hepatitis B infection. However, increased identification of HBV DNA positive but HBsAg negative cases has greatly promoted the use of molecular assays for more accurate HBV diagnosis. Over the past few decades, especially since the outbreak of COVID-19, significant advancements have been made in the techniques and devices for nucleic acid testing (NAT). Nowadays, the mainstream NAT techniques can broadly be split into two categories: PCR-based methods and non-PCR-based isothermal amplification methods. As achieving point-of-care testing (POCT) or on-site testing is an important development tendency for the next-generation NAT, non-PCR-based isothermal amplification methods like nucleic acid sequence-based amplification (NASBA), rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), helicase-dependent amplification (HDA), and recombinase polymerase amplification (RPA) have garnered significant attention in recent years. In this review, we provide a comprehensive overview of the nucleic acid isothermal amplification technologies currently used for HBV detection. The analytical performances of different methods are compared and their integration with microfluidics, lateral flow assays, and CRISPR/Cas systems is also discussed.

摘要

乙型肝炎病毒 (HBV) 感染是一个备受关注的全球性健康问题。临床上,血清学检测是最广泛应用的乙型肝炎病毒感染诊断检测方法,血清中 HBsAg 的存在表明存在急性和慢性乙型肝炎感染。然而,越来越多的 HBV DNA 阳性但 HBsAg 阴性病例的发现,极大地促进了更准确的乙型肝炎诊断的分子检测方法的应用。在过去几十年中,特别是自 COVID-19 爆发以来,核酸检测 (NAT) 的技术和设备取得了重大进展。如今,主流的 NAT 技术大致可分为两类:基于 PCR 的方法和非 PCR 恒温扩增方法。由于实现即时检测 (POCT) 或现场检测是下一代 NAT 的一个重要发展趋势,近年来,非 PCR 恒温扩增方法,如核酸序列扩增 (NASBA)、滚环扩增 (RCA)、环介导等温扩增 (LAMP)、解旋酶依赖性扩增 (HDA) 和重组酶聚合酶扩增 (RPA),受到了广泛关注。在这篇综述中,我们全面介绍了目前用于乙型肝炎病毒检测的核酸恒温扩增技术。比较了不同方法的分析性能,并讨论了它们与微流控、侧向流动检测和 CRISPR/Cas 系统的集成。

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