1. Rapid and Fully Microfluidic Ebola Virus Detection with CRISPR-Cas13a
- Author
-
Peiwu Qin, Myeongkee Park, Jean L. Patterson, Ricardo Carrion, Qian He, Manasi Tamhankar, Richard A. Mathies, Kendra J. Alfson, Ke Du, Ahmet Yildiz, and Anthony Griffiths
- Subjects
Diagnostic methods ,Microfluidics ,Bioengineering ,02 engineering and technology ,Biology ,medicine.disease_cause ,01 natural sciences ,Limit of Detection ,Lab-On-A-Chip Devices ,Fluorometer ,Endoribonucleases ,medicine ,CRISPR ,Fluorometry ,Instrumentation ,Leptotrichia ,Point of care ,Fluid Flow and Transfer Processes ,Ebola virus ,Base Sequence ,Process Chemistry and Technology ,010401 analytical chemistry ,Nucleic Acid Hybridization ,RNA ,Rna degradation ,Microfluidic Analytical Techniques ,Ebolavirus ,021001 nanoscience & nanotechnology ,Virology ,0104 chemical sciences ,Point-of-Care Testing ,RNA, Viral ,CRISPR-Cas Systems ,0210 nano-technology - Abstract
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 × 107 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.
- Published
- 2019