CRISPR检测技术应用于核酸现场即时检测系统综述

王雪琪; 何纳

doi:10.16462/j.cnki.zhjbkz.2024.11.016

CRISPR检测技术应用于核酸现场即时检测系统综述

doi: 10.16462/j.cnki.zhjbkz.2024.11.016

王雪琪,
何纳^,

复旦大学公共卫生学院流行病学教研室，上海 200032

基金项目:

国家重点研发计划项目 2022YFC2304902

上海市加强公共卫生体系建设三年行动计划重点学科建设项目 GWVI-11.1-05

详细信息

通讯作者:
何纳，E-mail: nhe@fudan.edu.cn

中图分类号: R183
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- PDF下载量: 19
- 被引次数: 0
出版历程
- 收稿日期: 2024-05-08
- 修回日期: 2024-08-19
- 网络出版日期: 2024-12-23
- 刊出日期: 2024-11-10

Systematic review of CRISPR applied to point-of-care nucleic acid detection

WANG Xueqi,
HE Na^,

Department of Epidemiology, School of Public Health, Fudan University, Shanghai 200032, China

Funds:

National Key Research and Development Program of China 2022YFC2304902

Key Disciplinary Program of Shanghai Public Health Action Plan GWVI-11.1-05

More Information

Corresponding author: HE Na, E-mail: nhe@fudan.edu.cn

摘要

摘要: 目的为了解成簇规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats, CRISPR)检测技术应用于核酸现场即时检测的研究现状。方法以“CRISPR”“nucleic acid detection”“point-of-care testing”或“POCT”为关键词，系统检索公开发表在PubMed、BioRxiv和MedRxiv数据库中的相关文献。结果根据纳入、排除标准，共纳入17篇文献，汇总分析CRISPR相关蛋白(CRISPR-associated protein, Cas)酶扩增方法、灵敏度、特异性、检测时间等方面信息。本研究结果显示，结合环介导等温扩增(loop-mediated isothermal amplification, LAMP)或重组酶聚合酶扩增(recombinase polymerase amplification, RPA)等核酸扩增方法后，CRISPR检测技术的灵敏度与实时荧光定量聚合酶反应(real time quantitative polymerase chain reaction, RT-qPCR)相当。结论 CRISPR检测过程在30~100 min即可完成，检测结果可肉眼读取，且检测成本与抗原检测相当，适用于资源匮乏地区进行大规模的核酸现场即时检测。
- 成簇规律间隔短回文重复序列 /
- 核酸检测 /
- 即时检测
Abstract: Objective To understand the current research status of clustered regularly interspaced short palindromic repeats (CRISPR) applied to nucleic acid detection. Methods In this study, we systematically reviewed related literatures published on databases of PubMed, BioRxiv, and MedRxiv using the keywords "CRISPR", "nucleic acid detection", "point-of-care testing" or "POCT". Results According to the inclusion and exclusion criteria, a total of 17 publications were selected. We summarized and analyzed the information of CRISPR-associated protein, amplification method, sensitivity, specificity, detection time and other aspects. Results suggested that when combined with nucleic acid amplification methods, such as loop-mediated isothermal amplification (LAMP) or recombinase polymerase amplification (RPA), the sensitivity of CRISPR was comparable to real time quantitative polymerase chain reaction (RT-qPCR). Conclusions The process of CRISPR can be completed within 30 to 100 minutes, with results that can be read by the naked eye. Additionally, the cost of detection is comparable to the antigen test, making it particularly suitable for large-scale point-of-care (POC) nucleic acid detection in resource limited areas.
- Clustered regularly interspaced short palindromic repeats /
- Nucleic acid detection /
- Point-of-care testing

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图 1 文献筛选流程图

Figure 1. Literature screening flowchart

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图 2 纳入文献的灵敏度和特异度森林图

Figure 2. Forest plots of sensitivity and specificity for included studies

下载: 全尺寸图片幻灯片

表 1 纳入文献的基本信息

Table 1. Basic information of the included publications

第一作者 First author	发表年份 Publication year	国家 Country	标本 Specimen	病原体 Pathogen	扩增方法 Amplification method	Cas酶 CRISPR-associated protein	时间 Time/min
Agrawal^[5]	2021	美国 United States	唾液 Saliva	SARS-CoV-2	LAMP	Cas13	35
Brandsma^[6]	2021	荷兰 Netherlands	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	LAMP	Cas12	30
Abugattas-Núñez^[7]	2024	秘鲁 Peru	唾液 Saliva	SARS-CoV-2	LAMP	Cas12a	40
Jirawannaporn^[8]	2022	泰国 Thailand	血清 Serum	钩端螺旋体 Leptospira	RPA	Cas12a	60
Ding^[9]	2021	中国 China	血清 Serum	乙型肝炎病毒 Hepatitis B virus	LAMP	Cas12	30 ~60
Qian^[10]	2021	中国 China	―	诺如病毒 Norovirus	RPA	Cas12a	40
Feng^[11]	2021	中国 China	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas12a	30
Wang^[12]	2021	中国 China	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	LAMP	Cas12a	45
Casati^[13]	2022	德国 Germany	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas13	60
Kham-Kjing^[14]	2022	泰国 Thailand	血浆 Plasma	丙型肝炎病毒 Hepatitis C virus	LAMP	Cas12	60 ~90
Patchsung^[15]	2023	泰国 Thailand	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas13	60
Lu^[16]	2022	中国 China	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas12a	20
Bhatt^[17]	2022	印度 India	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	LAMP	Cas12	< 100
Ali^[18]	2022	沙特阿拉伯 Saudi Arabia	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas9	< 60
Arizti-Sanz^[19]	2022	美国 United States	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	RPA	Cas13	< 90
Figueiredo^[20]	2023	葡萄牙 Portugal	鼻咽拭子 Nasopharyngeal swab	SARS-CoV-2	LAMP	Cas12a	40
Zhou^[21]	2023	中国 China	拭子 Swab	禽流感病毒 Avian influenza virus	RPA	Cas12a	60
注：1. SARS-CoV-2，严重急性呼吸综合征冠状病毒2型；LAMP，环介导等温扩增；RPA，重组酶聚合酶扩增。 2. “―”表示无法获取。 Note: 1. SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; LAMP, loop-mediated isothermal amplification; RPA, recombinase polymerase amplification. 2. "―" indicates that it cannot be obtained.

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