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To enable ultrasensitive and ultraspecific detection of single-base mutations in simple and economical ways, we have developed an artificial base mismatches-mediated PCR (ABM-PCR) detection approach.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2024-0962_abs_002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>ABM-PCR was applied to quantitative PCR (qPCR) and droplet digital PCR (ddPCR) detection platforms. The impact of mismatches on the thermodynamic stability of the primer-template duplex and the ability of Taq polymerase to catalyze the extension was examined. Effects of the sequence, position, and the number of mismatches on genotyping performance were characterized.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2024-0962_abs_003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>As proof of principle, we demonstrated the feasibility of ABM-PCR in detecting epidermal growth factor receptor (EGFR) and B-Raf proto-oncogene, serine\/threonine kinase (BRAF) mutations that are clinically relevant to diagnosis and prognosis of lung and thyroid cancers. Our ABM-PCR enabled the detection of 0.1\u202f% mutation without amplification of the wild-type DNA strand, even in the presence of a 300\u202fng human genomic DNA background. It enables ultrasensitive (\u226595\u202f%) and ultraspecific (\u226595\u202f%) diagnosis of clinical samples for thyroid papilloma and lung cancers. Based on these findings, we have established a set of rules and developed a user-friendly web primer design tool for designing effective ABM-PCR primers.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"j_cclm-2024-0962_abs_004\">\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This study highlights the impact of primer-template mismatches on PCR amplification and provides insights into rational design of effective ABM-PCR primers for detecting single-base mutations with high specificity and sensitivity. It is highly valuable for clinical diagnosis and prognosis use.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1515\/cclm-2024-0962","type":"journal-article","created":{"date-parts":[[2025,3,15]],"date-time":"2025-03-15T19:10:46Z","timestamp":1742065846000},"page":"1301-1314","source":"Crossref","is-referenced-by-count":5,"title":["Artificial base mismatches-mediated PCR (ABM-PCR) for detecting clinically relevant single-base mutations"],"prefix":"10.1515","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4528-0363","authenticated-orcid":false,"given":"Cia-Hin","family":"Lau","sequence":"first","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kejiang","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering , City University of Hong Kong , Hong Kong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Pathology , Fujian Cancer Hospital , Fuzhou , Fujian , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minghai","family":"Zou","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongqi","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Pediatrics , The Seventh Affiliated Hospital of Sun Yat-Sen University , Shenzhen , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhihao","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaqi","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenjiao","family":"Dong","sequence":"additional","affiliation":[{"name":"School of Public Health , Guangdong Medical University , Dongguan , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yumei","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pik Kwan","family":"Lo","sequence":"additional","affiliation":[{"name":"Department of Chemistry and State Key Laboratory of Marine Pollution , City University of Hong Kong , Hong Kong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongman","family":"Xue","sequence":"additional","affiliation":[{"name":"Department of Pediatrics , The Seventh Affiliated Hospital of Sun Yat-Sen University , Shenzhen , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojun","family":"Huang","sequence":"additional","affiliation":[{"name":"Xiamen Fly Gene Biomedical Technology CO., LTD, Biomedical Industrial Park , Xiamen , Fujian , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meijing","family":"Xu","sequence":"additional","affiliation":[{"name":"Xiamen Fly Gene Biomedical Technology CO., LTD, Biomedical Industrial Park , Xiamen , Fujian , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chung","family":"Tin","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering , City University of Hong Kong , Hong Kong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haibao","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Biology, College of Science , 12386 Shantou University , Shantou , Guangdong , China"},{"name":"Guangdong Provincial Key Laboratory of Marine Biotechnology , 12386 Shantou University , Shantou , Guangdong , China"},{"name":"Shantou Key Laboratory of Marine Microbial Resources and Interactions with Environment , 12386 Shantou University , Shantou , Guangdong , China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"374","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"2026051214225336869_j_cclm-2024-0962_ref_001","doi-asserted-by":"crossref","unstructured":"He, C, Wei, C, Wen, J, Chen, S, Chen, L, Wu, Y, et al.. 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