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Preamplification-free ultra-fast and ultra-sensitive point-of-care testing via LwaCas13a

Biosens Bioelectron. 2024-05;

Wanting Zeng, Wanping Chen, Yang Liu, Ting Zhang, Chao Zhai, Wenqiang Li, Longyu Wang, Cheng Zhang, Qili Zeng, Fei Wang, Lixin Ma

Products/Services Used	Details	Operation
Proteins, Expression, Isolation and Analysis	… The RNA used in this research were purchased from the GenScript Biotech Corporation (… et al., 2020), and three targets from the Nucleocapsid protein (N) gene fragment were selected …	Get A Quote

摘要

CRISPR based nucleic acid detection technology provides a deployable approach to point of care testing. While, there remain challenges limiting its practical applications, such as the need for pre-amplification and the long turnaround time. Here, we present a self-cascade signal amplification method with LwaCas13a and an artificially designed "U" rich RNA of stem-loop structure (URH) for pre-amplification-free ultra-fast and ultra-sensitive point-of-care testing (PASSPORT). The PASSPORT system contains: URH, crRNA targeted the URH, crRNA targeted the interesting RNA, fluorescent RNA reporter and LwaCas13a. The assay realized the detection of 100 copies/mL, within 5?min. The PASSPORT platform was further adopte... More