title: Sensitive and specific colorimetric DNA detection by invasive reaction coupled with nicking endonuclease-assisted nanoparticles amplification.
abstract: Colorimetric DNA detection is preferable to methods in clinical molecular diagnostics, because no expensive equipment is required. Although many gold nanoparticle-based colorimetric DNA detection strategies have been developed to analyze DNA sequences of interest, few of them can detect somatic mutations due to their insufficient specificity. In this study, we proposed a colorimetric DNA detection method by coupling invasive reaction with nicking endonuclease-assisted nanoparticles amplification (IR-NEANA). A target DNA firstly produces many flaps by invasive reaction. Then the flaps are converted to targets of nicking reaction-assisted nanoparticles amplification by ligation reaction to produce the color change of AuNPs, which can be observed by naked eyes. The detection limit of IR-NEANA was determined as 1pM. Most importantly, the specificity of the method is high enough to pick up as low as 1% mutant from a large amount of wild-type DNA backgrounds. The EGFR gene mutated at c.2573 T>G in 9 tissue samples from non-small cell lung cancer patients were successfully detected by using IR-NEANA, suggesting that our proposed method can be used to detect somatic mutations in biological samples.
authors.affiliation: Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Zou
authors.affiliation: Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Cao
authors.affiliation: Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China.
authors.forename: Haiping
authors.initials: H
authors.lastname: Wu
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Qinxin
authors.initials: Q
authors.lastname: Song
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Wang
authors.affiliation: Central Research Laboratory, Hitachi, Ltd., 1-280 Higashi-Koigakubo, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Kajiyama
authors.affiliation: Central Research Laboratory, Hitachi, Ltd., 1-280 Higashi-Koigakubo, Kokubunji-shi, Tokyo 185-8601, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Kambara
authors.affiliation: Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China; Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China; State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210009, China. Electronic address: ghzhou@nju.edu.cn.
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhou
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics; D003124:Colorimetry / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D018773:Genes, erbB-1; D006046:Gold / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D008175:Lung Neoplasms / Q000235:genetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods
chemical_list: D006046:Gold; D004247:DNA; D003850:Deoxyribonuclease I
keywords: AuNPs; Colorimetric detection; Invasive reaction; Ligation reaction; Mutation detection; Nicking endonuclease