HRP-integrated CRISPR-Cas12a biosensor for rapid point-of-care detection of Langya henipavirus
Xuan Wu,
Rong Xiang,
Danting Yang,
Xiaoyao He,
Lu Zhu,
Fangfang Sun,
Hanqing Li,
Ningning Pi,
Yi Li
Affiliations
Xuan Wu
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China; Hospital/School of Stomatology, Zunyi Medical University, 143 Dalian Road, Zunyi, Guizhou 563006, China; Hunan Provincial Key Laboratory of Medical Virology, College of Biology, Hunan University, 27 Tianma Road, Changsha, Hunan 410082, China
Rong Xiang
Precision Medicine Center, The Second Affiliated Hospital of Chongqing Medical University, 288 Tianwen Road, Chongqing 400010, China
Danting Yang
School of Public Health, Zhejiang Key Laboratory of Pathophysiology, Health Science Center, Ningbo University, 818 Fenghua Road, Ningbo, Zhejiang 315211, China
Xiaoyao He
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China
Lu Zhu
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China
Fangfang Sun
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China
Hanqing Li
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China
Ningning Pi
Key Laboratory of Environmental Chemistry and Ecotoxicology of Organic Pollutants of Chongqing, Ecological and Environment Monitoring Center of Chongqing, 252 Qishan Road, Chongqing 401132, China; Corresponding author
Yi Li
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), School of Laboratory Medicine, Chongqing Medical University, 1 Xueyuan Road, Chongqing 400016, China; Western (Chongqing) Collaborative Innovation Center for Intelligent Diagnostics and Digital Medicine, Chongqing National Biomedicine Industry Park, High-tech Zone, Chongqing 401329, China; Graduate School of Biomedical Engineering, Faculty of Engineering, ARC Centre of Excellence for Nanoscale Biophotonics, University of New South Wales, Sydney, NSW 2052, Australia; Corresponding author
Summary: Global pandemic has emphasized the needs for advanced pathogen diagnosis in dealing with newly emerged infectious threats, including the Langya henipavirus (LayV). LayV, as an emerging zoonotic pathogen, has potential to cause pandemic, but lacks of rapid diagnostic tools, particularly at point-of-care level. Here, we leveraged the merits of CRISPR-Cas12a biosensing and established a highly sensitive LayV detection method. By integrating CRISPR-Cas12a with RPA, 10 copies/μL ultra-sensitive LayV RNA detection has been achieved at room temperature within 30 min. More importantly, this study developed a special horseradish peroxidase (HRP)-single-stranded DNA (ssDNA) reporter enabling CRISPR-Cas12a to detect LayV RNA without pre-amplification, achieving a sensitivity of 1,200 copies/μL detectable by the naked eye. These explorations can serve as accelerator for CRISPR-Cas biosensing toward rapid response for newly emerged biological threats, and also provide a method to realize simple, precise, and amplicon-free point-of-care pathogen screening for resource limited or underdevelopment regions.
