Preamplification-free ultra-fast and ultra-sensitive point-of-care testing via LwaCas13a.
Biosens Bioelectron
; 259: 116400, 2024 Sep 01.
Article
em En
| MEDLINE
| ID: mdl-38776799
ABSTRACT
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 adopted for the detection of marker gene from SASR-CoV-2 and Severe fever with thrombocytopenia syndrome virus (SFTSV), respectively, and 100% accuracy for the analysis of clinical specimens (100 SASR-CoV-2 specimens and 16 SFTSV specimens) was obtained. Integrated with a lateral flow assay device, this assay could provide an alternative platform for the development of point of care testing (POCT) biosensors. PASSPORT has the potential to enable sensitive, specific, user-friendly, rapid, affordable, equipment-free and point-of-care testing for the purpose of large-scale screening and in case of epidemic outbreak.
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MEDLINE
Assunto principal:
Técnicas Biossensoriais
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Sistemas CRISPR-Cas
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Testes Imediatos
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SARS-CoV-2
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COVID-19
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article