Cong Wang1, Xiaomin Chen1, Yuying Wu1, Hao Li1, Yu Wang1, Xiaofu Pan2, Tingting Tang2, Ziying Liu1, Xiaokun Li3. 1. School of Pharmacy, Wenzhou Medical University, Wenzhou, 325030, Zhejiang, China. 2. The Third Affiliated Hospital of Wenzhou Medical University, Ruian, 325200, Zhejiang, China. 3. School of Pharmacy, Wenzhou Medical University, Wenzhou, 325030, Zhejiang, China. xiaokunli@163.net.
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.
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.
Authors: Panagiota M Kalligosfyri; Sofia Nikou; Sofia Karteri; Haralabos P Kalofonos; Vasiliki Bravou; Despina P Kalogianni Journal: Biosensors (Basel) Date: 2022-02-04