Abstract
Objectives
To develop a convenient and sensitive point-of-care test for detecting gene mutations based on allele-specific PCR.
Results
To develop a lateral flow strip for visual detection of K-ras mutations based on a modified PCR, a specific DNA tag was covalently linked to the 5′-end of each primer by a nine-carbon linker to produce a sticky end. One of the sticky ends of the PCR products bound to gold nano-particles, while the other sticky end was captured onto a nitrocellulose membrane of lateral flow strips. The lateral flow strip showed a great sensitivity, which detected mutations in as low as 10 tumor cells. The positive rate and accuracy of the lateral flow strip for blood samples were over 92 and 96 %, respectively.
Conclusions
The lateral flow strip provides an easy method for sensitive detection of gene mutations based on allele specific-PCR.
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Acknowledgments
This work was supported by the Natural Science Foundation of Zhejiang Province of China (LY16H140004, Y2110492) to CW, the National Natural Science Foundation of China (81101712, 81270761, 31371470) to CW, the Natural Science Foundation of Ningbo city (2015A610198), and the Talent project foundation of Wenzhou Medical University (QTJ14030).
Supporting information
Supplementary Table 1—Primer sequence.
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Cong Wang, Xiaomin Chen, Yuying Wu have equally contributed to this work.
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Wang, C., Chen, X., Wu, Y. et al. Lateral flow strip for visual detection of K-ras mutations based on allele-specific PCR. Biotechnol Lett 38, 1709–1714 (2016). https://doi.org/10.1007/s10529-016-2161-9
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DOI: https://doi.org/10.1007/s10529-016-2161-9