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清華程京院士課題組采用我司硅羧基磁珠PMSi300003提取咽拭子檢測(cè)新冠

Sensitive and Rapid Diagnosis of Respiratory Virus Coinfection Using a Microfluidic Chip-Powered CRISPR/Cas12a System

使用微流控芯片驅(qū)動(dòng)的 CRISPR/Cas12a 系統(tǒng)靈敏快速地診斷呼吸道病毒感染
Jiajia Liu,Huili Wang,Li Zhang,Ying Lu,Xu Wang,Minjie Shen,Nan Li,Li Feng,Juhui Jing,Bin Cao,Xiaohui Zou,Jing Cheng,Youchun Xu
First published: 22 May 2022 https://doi.org/10.1002/smll.202200854


Abstract
The ongoing pandemic caused by severe acute respiratory syndrome coronavirus 2 is profoundly influencing the global healthcare system and people's daily lives. The high resource consumption of coronavirus disease 2019 (COVID-19) is resulting in insufficient surveillance of coinfection or resurgence of other critical respiratory epidemics, which is of public concern. To facilitate evaluation of the current coinfection situation, a microfluidic system (MAPnavi) is developed for the rapid (<40?min) and sensitive diagnosis of multiple respiratory viruses from swab samples in a fully sealed and automated manner, in which a nested-recombinase polymerase amplification and the CRISPR-based amplification system is first proposed to ensure the sensitivity and specificity. This novel system has a remarkably low limit of detection (50–200 copies mL?1) and is successfully applied to detect 171 clinical samples (98.5% positive predictive agreement; 100% negative predictive agreement), and the results identify 45.6% coinfection among clinical samples from patients with COVID-19. This approach has the potential to shift diagnostic and surveillance efforts from targeted testing for a high-priority virus to comprehensive testing of multiple virus sets and to greatly benefit the implementation of decentralized testing.

摘要 

      由嚴(yán)重急性呼吸綜合征冠狀病毒 2 引起的持續(xù)大流行正在深刻影響全球醫(yī)療保健系統(tǒng)和人們的日常生活。 2019 年冠狀病毒病 (COVID-19) 的高資源消耗導(dǎo)致對(duì)合并感染或其他嚴(yán)重呼吸道傳染病復(fù)發(fā)的監(jiān)測(cè)不足，這是公眾關(guān)注的問(wèn)題。為了便于評(píng)估當(dāng)前的共感染情況，開(kāi)發(fā)了一種微流控系統(tǒng) (MAPnavi)，用于以完全密封和自動(dòng)化的方式快速（<40?min）和靈敏地診斷拭子樣本中的多種呼吸道病毒，其中嵌套重組酶聚合酶首次提出了基于CRISPR的擴(kuò)增系統(tǒng)，以確保靈敏度和特異性。這種新型系統(tǒng)具有非常低的檢測(cè)限（50-200 拷貝 mL-1），并成功應(yīng)用于檢測(cè) 171 個(gè)臨床樣本（98.5% 的陽(yáng)性預(yù)測(cè)一致性；100% 的陰性預(yù)測(cè)一致性），結(jié)果確定了 45.6% 的共感染來(lái)自 COVID-19 患者的臨床樣本。這種方法有可能將診斷和監(jiān)測(cè)工作從針對(duì)高優(yōu)先級(jí)病毒的針對(duì)性測(cè)試轉(zhuǎn)變?yōu)閷?duì)多個(gè)病毒集的全面測(cè)試，并極大地有利于分散測(cè)試的實(shí)施。

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