title: A universal probe design for colorimetric detection of single-nucleotide variation with visible readout and high specificity.
abstract: Single-nucleotide variation (SNV) is a crucial biomarker for drug resistance-related detection in cancer and bacterial infection. However, the unintended binding of DNA probes limits the specificity of SNV detection, and the need for redesigned sequences compromise the universality of SNV assay. Herein, we demonstrated a universal and low-cost assay for the colorimetric discrimination of drug-resistance related point mutation. By the use of a universal DNA probe and a split G-quadruplex, the signal could be recognized by naked eye at room temperature. The DNA probe was used as a signal reporter which not only improved the universality, but also enabled high specificity of probe hybridization. This assay was successfully applied in the detection of cancer-related SNV in the epidermal growth factor receptor (EGFR) gene, kirsten rat sarcoma viral oncogene homologue (KRAS), and tuberculosis drug-resistance related point mutation in RNA polymerase beta subunit gene (rpoB) with high specificity and visible readout. This method was simple, rapid, high-throughput and effective, which was suitable for point-of-care applications.
authors.affiliation: Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Xueping
authors.initials: X
authors.lastname: Chen
authors.affiliation: Clinical Laboratories, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Huawei
authors.initials: H
authors.lastname: Shen
authors.affiliation: Clinical Laboratories, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Wenli
authors.initials: W
authors.lastname: Feng
authors.affiliation: Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, P. R. China.
authors.forename: Guoming
authors.initials: G
authors.lastname: Xie
mesh_terms: D003124:Colorimetry / Q000379:methods* / Q000592:standards; D015342:DNA Probes; D066246:ErbB Receptors / Q000235:genetics; D054856:G-Quadruplexes; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D066246:ErbB Receptors; D016283:Proto-Oncogene Proteins p21(ras)
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