RNA-based isothermal amplification technology and its clinical application in pathogen infection

IF 3.6 3区 化学 Q2 CHEMISTRY, ANALYTICAL Reviews in Analytical Chemistry Pub Date : 2022-01-01 DOI:10.1515/revac-2022-0051
Jie Teng, Fang Liu, Li Chang, Qiu-xia Yang, Guanglu Che, Shuyu Lai, Yuan Tan, Jiaxin Duan, Yongmei Jiang
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Abstract

Abstract It is very important to detect pathogenic bacteria, viruses, or fungi in a patient’s secretion or body fluid samples as soon as possible to determine the patient’s recovery. For certain pathogens, the amount of ribosomal RNA copies contained is often tens of thousands of times higher than the amount of DNA copies, so the detection of RNA has higher sensitivity. In addition, whether for DNA pathogens or RNA pathogens, the direct detection of ribonucleic acid transcribed by pathogens in vivo can distinguish active infection or past infection, can eliminate the influence of residual DNA of pathogens that have died in the lesions, and can also avoid excessive medical interventions for transient infections, which is of great significance in the field of infectious pathogen detection. Isothermal amplification technology played important roles in molecular diagnosis because of its significant advantages. Highly sensitive RNA detection can be achieved by both direct transcription amplification and indirect amplification based on reverse transcription. Direct transcription amplification technologies relies on reverse transcriptase and T7 RNA polymerase to achieve linear transcription amplification of RNA on one-step; while the indirect amplification technology depends on a reverse transcriptional process at the beginning of the reaction. Both methods have outstanding advantages in clinical application, and commercial kits and commercial all-in-one machines based on these principles have been put into clinical use. This review mainly introduces the clinical application of isothermal amplification technologies in the detection of RNA pathogens and the main difficulties faced at this stage. It is hoped to provide insightful ideas for the construction of pathogen RNA detection technology to meet the needs of point-of-care testing in the future.
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基于rna的等温扩增技术及其在病原体感染中的临床应用
摘要尽早检测患者分泌物或体液样本中的致病菌、病毒或真菌,对判断患者的康复情况非常重要。对于某些病原体,核糖体所含RNA拷贝数往往比DNA拷贝数高数万倍,因此对RNA的检测具有更高的灵敏度。此外,无论是DNA病原体还是RNA病原体,直接检测病原体在体内转录的核糖核酸,可以区分活动性感染还是既往感染,可以消除病变中已死亡病原体残留DNA的影响,也可以避免对暂时性感染的过度医疗干预,在感染性病原体检测领域具有重要意义。等温扩增技术以其显著的优势在分子诊断中发挥着重要的作用。通过直接转录扩增和基于逆转录的间接扩增均可实现高灵敏度的RNA检测。直接转录扩增技术依靠逆转录酶和T7 RNA聚合酶一步实现RNA的线性转录扩增;而间接扩增技术依赖于反应开始时的逆转录过程。这两种方法在临床应用中都有突出的优势,基于这些原理的商用套件和商用一体机已经投入临床使用。本文主要介绍了等温扩增技术在RNA病原体检测中的临床应用及现阶段面临的主要困难。希望能为今后病原体RNA检测技术的构建提供有见地的思路,以满足护理点检测的需求。
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来源期刊
Reviews in Analytical Chemistry
Reviews in Analytical Chemistry 化学-分析化学
CiteScore
7.50
自引率
0.00%
发文量
15
审稿时长
>12 weeks
期刊介绍: Reviews in Analytical Chemistry publishes authoritative reviews by leading experts in the dynamic field of chemical analysis. The subjects can encompass all branches of modern analytical chemistry such as spectroscopy, chromatography, mass spectrometry, electrochemistry and trace analysis and their applications to areas such as environmental control, pharmaceutical industry, automation and other relevant areas. Review articles bring the expert up to date in a concise manner and provide researchers an overview of new techniques and methods.
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