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: Xueping Chen, Dandan Zhou, Huawei Shen, Hui Chen, Wenli Feng, Guoming Xie
• Journal: Scientific reports (Sci Rep) Vol. 6 Pg. 20257 (Feb 02 2016) ISSN: 2045-2322 [Electronic] England
• PMID: 26830326 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA Probes
• ErbB Receptors
• Proto-Oncogene Proteins p21(ras)

Topics

• Colorimetry (methods, standards)
• DNA Probes
• ErbB Receptors (genetics)
• G-Quadruplexes
• Humans
• Polymorphism, Single Nucleotide
• Proto-Oncogene Proteins p21(ras) (genetics)
• Sensitivity and Specificity