A low-cost novel lateral flow nucleic acid assay (LFNAA) for yak milk authentication

文献类型： 外文期刊

第一作者： Wang, Zhiying

作者： Wang, Zhiying;Li, Tingting;Yu, Wenjie;Qiao, Lu;Yang, Shuming;Chen, Ailiang

作者机构：

关键词： Recombinant polymerase amplification (RPA); Lateral flow nucleic acid assay (LFNAA); Spacer C3; ssDNA tail; Yak milk authentication

期刊名称：LWT-FOOD SCIENCE AND TECHNOLOGY （ 影响因子：4.952； 五年影响因子：5.383 ）

ISSN： 0023-6438

年卷期： 2020 年 122 卷

页码：

收录情况： SCI

摘要： A low-cost novel lateral flow nucleic acid assay (LFNAA) was developed for rapid and sensitive authentication of yak milk on-site. In this method, a special abasic spacer C3 and tail nucleotide sequence modified reverse primers were innovatively designed for rapid isothermal recombinant polymerase amplification (RPA) to obtain a double-stranded amplicon with a single-stranded DNA tail, which could directly bind to gold nanoparticle (AuNP)-labeled detection probes, and produce a sandwich complex to show a red color on the test line of the strip. Compared to conventional RPA-lateral flow assay based on expensive antibodies, the proposed method in this work demonstrated massive cost reduction, avoided time-consuming and laborious AuNP-antibody conjugation process. The LFNAA revealed high specificity and sensitivity, which allowed detecting as low as 5% of yak milk within 40 min. This LFNAA facilitated non-specialized operators to field determine the authenticity of yak milk by naked eyes, and could be readily applied to other nucleic acid detection such as food safety inspection and point-of-care medical tests.

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