Rapid and Fully Microfluidic Ebola Virus Detection with CRISPR-Cas13a

被引：249

作者：

Qin, Peiwu ^{[1
,2
]}

Park, Myeongkee ^{[3
,4
]}

Alfson, Kendra J. ^{[5
]}

Tamhankar, Manasi ^{[5
]}

Carrion, Ricardo ^{[5
]}

Patterson, Jean L. ^{[5
]}

Griffiths, Anthony ^{[5
,6
,7
]}

He, Qian ^{[2
,8
]}

Yildiz, Ahmet ^{[1
]}

Mathies, Richard ^{[3
]}

Du, Ke ^{[3
,8
,9
]}

机构：

[1] Univ Calif Berkeley, Dept Phys, Berkeley, CA 94720 USA

[2] Tsinghua Berkeley Shenzhen Inst, Ctr Precis Med & Healthcare, Shenzhen 518055, Guangdong, Peoples R China

[3] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

[4] Dong A Univ, Dept Chem, Busan 49315, South Korea

[5] Texas Biomed Res Inst, Dept Virol & Immunol, San Antonio, TX 78227 USA

[6] Boston Univ, Dept Microbiol, Boston, MA 02118 USA

[7] Boston Univ, Natl Emerging Infect Dis Labs, Boston, MA 02118 USA

[8] Rochester Inst Technol, Dept Mech Engn, Rochester, NY 14623 USA

[9] Rochester Inst Technol, Dept Microsyst Engn, Rochester, NY 14623 USA

来源：

ACS SENSORS | 2019年 / 4卷 / 04期

关键词：

CRISPR; Ebola; microfluidics; amplification free; fluorescence; point-of-care; NUCLEIC-ACID DETECTION; AMPLIFICATION-FREE; WEST-AFRICA; RT-PCR; ASSAY; QUANTIFICATION; PLATFORM; POINT;

D O I：

10.1021/acssensors.9b00239

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Highly infectious illness caused by pathogens is endemic especially in developing nations where there is limited laboratory infrastructure and trained personnel. Rapid point-of-care (POC) serological assays with minimal sample manipulation and low cost are desired in clinical practice. In this study, we report an automated POC system for Ebola RNA detection with RNA-guided RNA endonuclease Cas13a, utilizing its collateral RNA degradation after its activation. After automated microfluidic mixing and hybridization, nonspecific cleavage products of Cas13a are immediately measured by a custom integrated fluorometer which is small in size and convenient for in-field diagnosis. Within 5 min, a detection limit of 20 pfu/mL (5.45 X 10(7) copies/mL) of purified Ebola RNA is achieved. This isothermal and fully solution-based diagnostic method is rapid, amplification-free, simple, and sensitive, thus establishing a key technology toward a useful POC diagnostic platform.

引用

页码：1048 / 1054

页数：13

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