Details

Autor(en) / Beteiligte

• He, Qian
• Yu, Dongmei
• Bao, Mengdi
• Korensky, Grant
• Chen, Juhong
• Shin, Mingyeong
• Kim, Juwon
• Park, Myeongkee
• Qin, Peiwu
• Du, Ke

Titel

High-throughput and all-solution phase African Swine Fever Virus (ASFV) detection using CRISPR-Cas12a and fluorescence based point-of-care system

Ist Teil von

• Biosensors & bioelectronics, 2020-04, Vol.154, p.112068-112068, Article 112068

Ort / Verlag

England: Elsevier B.V

Erscheinungsjahr

2020

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Here we report the development of a high throughput, all-solution phase, and isothermal detection system for African Swine Fever Virus (ASFV). CRISPR-Cas12a programmed with a CRISPR RNA (crRNA) is used to detect ASFV target DNA. Upon ASFV DNA binding, the Cas12a/crRNA/ASFV DNA complex becomes activated and degrades a fluorescent single stranded DNA (ssDNA) reporter present in the assay. We combine this powerful CRISPR-Cas assay with a fluorescence-based point-of-care (POC) system for rapid and accurate virus detection. Without nucleic acid amplification, a detection limit of 1 pM is achieved within 2 h. In addition, the ternary Cas12a/crRNA/ASFV DNA complex is highly stable at physiological temperature and continues to cleave the ssDNA reporter even after 24 h of incubation, resulting in an improved detection limit of 100 fM. We show that this system is very specific and can differentiate nucleic acid targets with closely matched sequences. The high sensitivity and selectivity of our system enables the detection of ASFV in femtomolar range. Importantly, this system features a disposable cartridge and a sensitive custom designed fluorometer, enabling compact and simple ASFV detection, intended for low resource settings. •A point-of-care device has been developed for rapid and sensitive African Swine Fever Virus detection.•This system combines an all-solution phase CRISPR assay with a custom designed fluorescence sensing unit.•Without target amplification, a detection limit of 1 pM is reported within 2 hrs.