title: A colorimetric method for H1N1 DNA detection using rolling circle amplification.
abstract: A highly sensitive and specific colorimetry-based rolling circle amplification (RCA) assay has been successfully developed as a method for the effective detection of H1N1 DNA. Specific oligonucleotide and reporter primer probes were designed together with a circular template, and the oligonucleotide probes were attached to the surfaces of magnetic beads (MBs) to form functional MB-DNA conjugates as capture probes for the target H1N1 DNA molecules. Together with the addition of DNA targets and reporter primer probes to the MB-DNA conjugates, sandwiched hybrids were formed. The initiation of RCA amplification using the circular template in the presence of phi29 polymerase allowed for the amplification of a large number of repeat sequences of the single-stranded (ss)-DNA product. This RCA product accumulated gold nanoparticles (AuNPs), resulting in a colorimetric change that could be viewed by the naked eye or detected using UV-vis spectroscopy. According to this method, H1N1 DNA could be detected at the 1 pmol L(-1) level. This platform exhibited design convenience, simplicity, and cost-effectiveness, and could be used to provide a new diagnostic assay for H1N1, and other infectious diseases.
authors: State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai, 200050, China.
authors: Yasi
authors: Y
authors: Xing
authors: Ping
authors: P
authors: Wang
authors: Yucui
authors: Y
authors: Zang
authors: Yuqing
authors: Y
authors: Ge
authors: Qinghui
authors: Q
authors: Jin
authors: Jianlong
authors: J
authors: Zhao
authors: Xia
authors: X
authors: Xu
authors: Guoqiang
authors: G
authors: Zhao
authors: Hongju
authors: H
authors: Mao
mesh_terms: D003124:Colorimetry / Q000379:methods*; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics*; D005240:Feasibility Studies; D006046:Gold / Q000737:chemistry; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*
chemical_list: D004279:DNA, Viral; D006046:Gold