CRISPR-cas12a mediated SERS lateral flow assay for amplification-free detection of double-stranded DNA and single-base mutation

Yuanfeng Pang*, Qing Li, Chongwen Wang, Shuai zhen, Zhiwei Sun, Rui Xiao

*此作品的通讯作者

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85 引用 (Scopus)

摘要

Lateral flow assay (LFA) is user-friendly diagnostic tools but suffering limitations in poorer sensitivities and specificities especially for double-stranded DNA and single-base mutation quantification. Here, to improve the sensitivity and specificity for the LFA based nucleic acid detection, CRISPR-Cas12a mediated Surface enhanced Raman scattering (SERS) LFA was developed. By combination of ultra-sensitive SERS tags and target-specific signal amplification ability of CRISPR-Cas12a, HIV-1 dsDNA can be directly quantified with a LOD of 0.3 fM without any pre-amplification steps, which is almost 4 orders of magnitude lower than that of traditional colorimetric LFA methods. The whole detection process can be finished less than 1 h. Moreover, based on the target specificity of Cas12a, HIV-1 single-based drug resistance mutation (M184V) can be recognized as low as 0.01%. The HIV-1 dsDNA can also be successfully detected in serum samples with good comparable of that in buffer setting. Therefore, the simple and inexpensive paper-based CRISPR-SERS strip has great potential for point-of-care testing (POCT) of nucleic acid targets especially in resource-poor or non-laboratory environments.

源语言英语
文章编号132109
期刊Chemical Engineering Journal
429
DOI
出版状态已出版 - 1 2月 2022
已对外发布

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