A micro chip for rapid detection of multiple types of respiratory virus.
Paper:
CRISPR/Cas12a-Assisted isothermal amplification for rapid and specific diagnosis of respiratory virus on an microfluidic platform
Abstract
have long been a major cause of a global pandemic, emphasizing the urgent need for high-sensitivity diagnostic tools. Typical PCR technology can only determine the type of virus in the sample, which is unable to detect different variants of the same virus without costly and time-consuming gene sequencing. Here, we introduce a simple, fully enclosed, and highly integrated
system based on CRISPR/Cas12a and
amplification techniques (LOC-CRISPR) that can specifically identify multiple common respiratory viruses and their variants. The LOC-CRISPR chip integrates viral
extraction, recombinant
amplification, and CRISPR/Cas12a cleavage reaction-based detection, contamination-free detection. In addition, the LOC-CRISPR chip was designed for multiplexed detection (two-sample input and ten-result outputs), which can not only detect the presence of SARS-CoV-2, H1N1, H3N2, IVB and
but also differentiate the BA.1, BA.2, and BA.5 variants of SARS-COV-2. For clinical validation, the LOC-CRISPR chip was used to analyze 50 nasopharyngeal swab samples (44 positive and 6 negative) and achieved excellent sensitivity (97.8%) and specificity (100%). This innovative LOC-CRISPR system has the ability to quickly, sensitively, and accurately detect multiple target
sequences with single-base mutations, which will further improve the rapid identification and traceability of respiratory viruses infectious diseases.
News release:
中国科学院深圳先进技术研究院生物医学与健康工程研究所杨慧研究员团队,与深圳市儿童医院麻晓鹏、谢中建团队合作,在
Biosensors and Bioelectronics上发表了题为
CRISPR/Cas12a-Assisted isothermal amplification for rapid and specific diagnosis of respiratory virus on an microfluidic platform的文章。该研究开发了一种基于CRISPR/Cas12a和等温扩增技术(LOC-CRISPR)的简单、全封闭、高度集成的微流控系统,可以特异性识别多种常见呼吸道病毒及其变种。本研究中,沈杰男博士、陈挚博士和谢瑞彬为共同第一作者,麻晓鹏院长、杨慧研究员和谢中建教授为共同通讯作者,深圳市儿童医院与中国科学院深圳先进技术研究院为通讯单位。
呼吸道病毒是导致呼吸道感染的主要因素之一,包括冠状病毒、流感病毒和呼吸道合胞病毒(RSV)等。在儿童和老年人等高风险人群中,病毒感染导致的死亡率极高。传统的诊断方法通常需要复杂的实验设备和较长的检测周期,且存在着高成本、低灵敏度等问题。并且传统的PCR技术只能确定病毒类型,如果要检测同一病毒的不同变异株,就需要进行昂贵且耗时的基因测序。为了解决上述问题,研究人员开发了一种名为LOC-CRISPR的新技术,它基于CRISPR/Cas12a和等温扩增技术,是一种简单、全封闭、高度集成的微流控系统。该系统可以对SARS-CoV-2(新冠)及其变异株(BA.1、BA.2和BA.5)、H1N1(甲型流感病毒)、H3N2(甲型流感病毒)、IVB(流感病毒)和HRSV(呼吸道感染病毒)的进行精准识别。在临床样本测试中,研究团队使用LOC-CRISPR芯片分析了50份鼻咽拭子样本(44份阳性和6份阴性),并获得了优异的灵敏度(97.8%)和特异性(100%)。
该系统中,研究人员在LOC-CRISPR芯片中集成了核酸提取、核酸扩增和荧光检测三个功能区。并且LOC-CRISPR芯片还设计了用于多重检测(两个样本输入和十个结果输出)需求的特殊结构。该系统中的另一项关键技术是CRISPR/Cas12a技术,它是一种基于RNA导向的基因编辑技术,能够通过特异性识别和切割DNA序列,具有高度特异性和高效率的特点。研究团队利用CRISPR/Cas12a的导向作用,将其与等温扩增技术相结合,减少检测步骤,降低检测时间,不仅实现了呼吸道病毒的种类识别,而且还实现了对新型冠状病毒的不同变异株之间的碱基序列识别。
总之,该项研究的进展,具有将实验室科学带入社区诊所的能力,是诊断技术领域的一个重大突破。未来,随着针对多种疾病诊断试剂的持续开发,该款微流控芯片将在疾病预防、健康监测以及个性化医疗方面,具有巨大的潜力和应用价值,值得各方关注和期待。