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可视化核酸杂交侧流条与微针集成用于即时护理认证

Visualized Nucleic Acid Hybridization Lateral Flow Strip Integrating with Microneedle for the Point-of-Care Authentication of .

作者信息

Liu Haibin, Wang Xinyue, Tian Hang, Yuan Yi, Wang Jing, Cheng Yani, Sun Linyao, Chen Hongshuo, Song Xiaoming

机构信息

College of Life Sciences, North China University of Science and Technology, Tangshan 063200, China.

College of Electrical Engineering, North China University of Science and Technology, Tangshan 063200, China.

出版信息

Int J Mol Sci. 2024 Dec 19;25(24):13599. doi: 10.3390/ijms252413599.

DOI:10.3390/ijms252413599
PMID:39769360
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11677120/
Abstract

Due to the price and demand of having increased dramatically, adulteration with other fungi is a common problem. Thus, a reliable method of authentic identification is essential. In the present work, a rapid DNA extraction and double-tailed recombinase polymerase amplification (RPA) coupled with nucleic acid hybridization lateral flow strip (NAH-LFS) was developed to distinguish authentic ingredients from other fungi substitutes. In the presence of , the RPA amplicons with two ssDNA tails in the opposite ends, which could simultaneously bind with the SH-probes on gold nanoparticles (AuNPs) and capture the probe on the test line, formed visible red bands. RPA combined with NAH-LFS can efficiently detect DNA down to 1.4 ng/μL; meanwhile, the specificity test validated no cross reaction with common adulterants, including , , , , and . The whole RPA-NAH-LFS could be completed within 16 min. The RPA-NAH-LFS results in detecting 20 commercial samples are consistent with PCR-AGE and RT-PCR, confirming the feasibility of the RPA-NAH-LFS method. In conclusion, these results are expected to facilitate the application of RPA-NAH-LFS in the authentication detection of materials, providing a convenient and efficient method for quality control.

摘要

由于价格和需求急剧增加，与其他真菌掺假是一个常见问题。因此，可靠的正品鉴定方法至关重要。在本研究中，开发了一种快速DNA提取方法，并结合双尾重组酶聚合酶扩增（RPA）与核酸杂交侧流试纸条（NAH-LFS），以区分正品成分与其他真菌替代品。在存在[具体物质]的情况下，两端带有两条单链DNA尾巴的RPA扩增子，可同时与金纳米颗粒（AuNPs）上的SH探针结合，并与检测线上的捕获探针结合，形成可见的红色条带。RPA与NAH-LFS相结合能够有效检测低至1.4 ng/μL的[具体物质]DNA；同时，特异性测试验证了与常见掺假物（包括[列举的掺假物]）无交叉反应。整个RPA-NAH-LFS检测过程可在16分钟内完成。RPA-NAH-LFS检测20个商业[具体物质]样品的结果与PCR-AGE和RT-PCR一致，证实了RPA-NAH-LFS方法的可行性。总之，这些结果有望促进RPA-NAH-LFS在[具体物质]材料真伪检测中的应用，为[具体物质]质量控制提供一种便捷高效的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/32fdee973c5a/ijms-25-13599-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/792ec293c666/ijms-25-13599-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/d6f932fef50f/ijms-25-13599-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/03add38ddb09/ijms-25-13599-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/b47cbfc79f76/ijms-25-13599-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/e65ecf6996f3/ijms-25-13599-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/a39e4f326627/ijms-25-13599-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/0c21f355cf16/ijms-25-13599-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/7cd7b23d338f/ijms-25-13599-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/5c5380183668/ijms-25-13599-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/32fdee973c5a/ijms-25-13599-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/792ec293c666/ijms-25-13599-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/d6f932fef50f/ijms-25-13599-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/03add38ddb09/ijms-25-13599-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/b47cbfc79f76/ijms-25-13599-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/e65ecf6996f3/ijms-25-13599-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/a39e4f326627/ijms-25-13599-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/0c21f355cf16/ijms-25-13599-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/7cd7b23d338f/ijms-25-13599-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/5c5380183668/ijms-25-13599-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa7f/11677120/32fdee973c5a/ijms-25-13599-g010.jpg

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