基于CRISPR-Cas13a/Cas12a的快速灵敏地检测猴痘病毒及其共感染病毒的双靶标一步检测方法  

Rapid and sensitive Cas13a/Cas12a-based one-pot dual-target strategy to detect monkeypox virus and its co-infected viruses

作　　者：王潇 饶巧 陆正荣 邓小宝 沈容 王瑞 董玮艺 杞仙花 靳珍 汤有志 杜丹 Xiao Wang;Qiao Rao;Zhengrong Lu;Xiaobao Deng;Rong Shen;Rui Wang;Weiyi Dong;Xianhua Qi;Zhen Jin;Youzhi Tang;Dan Du(State Key Laboratory of Cellular Stress Biology,Cacner Research Center,School of Medicine,Faculty of Medicine and Life Sciences,Xiamen University,Xiamen 361102,China;Department of Stomatology,School of Medicine,Xiamen University,Xiamen 361102,China;Innovation Center for Cell Signaling Network,Xiamen University,Xiamen 361102,China;Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation,College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]State Key Laboratory of Cellular Stress Biology,Cacner Research Center,School of Medicine,Faculty of Medicine and Life Sciences,Xiamen University,Xiamen 361102,China [2]Department of Stomatology,School of Medicine,Xiamen University,Xiamen 361102,China [3]Innovation Center for Cell Signaling Network,Xiamen University,Xiamen 361102,China [4]Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation,College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China

出　　处：《Science Bulletin》2023年第24期3142-3148,M0005,共8页科学通报（英文版）

基　　金：supported by the National Key R&D Program of China(2022YFA1304000);the National Natural Science Foundation of China(81972823);the Foundation for Innovative Research Groups of the National Natural Science Foundation of China(32121004);the Guangdong Special Support Program innovation team(2019BT02N054);the Open Research Fund of State Key Laboratory of Cellular Stress Biology,Xiamen University(SKLCSB2019KF013);the Fundamental Research Funds for the Central University(20720220003)。

摘　　要：近期全球范围内的猴痘病毒(MPXV)爆发引发了与其他病原体的严重共感染,尤其在资源匮乏的地区,急需快速而精准的现场诊断方案.该研究基于CRISPR-Cas13a和Cas12a系统的反式切割特性,分别建立了单靶标的一步检测方法,并整合这两个系统建立了双重靶标检测系统.Cas13a和Cas12a的单靶标一步检测方法展现出高于标准定量PCR的阳性检测率,检测极限达到4拷贝,总检测时间不超过40min,而且无需复杂设备或专业技术培训.而基于Cas13a/Cas12a的双重靶标检测系统,可检测到低至阿摩尔浓度的靶标,总检测时间不超过45 min.这一双靶标一步检测方法可以用于多靶标鉴定MPXV,能够在单一样本中区分MPXV亚型,或鉴定MPXV与SARS-CoV-2或HIV病毒的共感染情况.经过临床模拟样本的检验,结果确认了这种方法的快速、准确、可靠、方便且高效的特性,凸显了其应用于现场检测方面的潜力,有助于改进疾病检测方法和传染病管理.Monkeypox(MPX)is a zoonotic infection recognized as an emergent disease and a global threat by the World Health Organization(WHO)and the Centers for Disease Control and Prevention(CDC)[1].MPXV is transmitted through contact with infected animals or through direct contact with the body fluids,blood,respiratory droplets,lesions,or contaminated personal objects of natural hosts or patients.

关 键 词：共感染 专业技术培训 疾病检测 传染病管理 现场诊断 检测极限 猴痘病毒 摩尔浓度 

分 类 号：R511[医药卫生—内科学]