使用金纳米粒子比色法检测环介导等温扩增的DNA.docVIP

  Colorimetric detection of amplified target DNA by loop-mediated isothermal amplification using gold 5 10 15 20 nanoparticle# Abstract: We have developed a rapid, simple, and label-free colorimetric method for the identification of target DNA. It is based on loop-mediated isothermal amplification (LAMP). Plain gold nanoparticles (AuNPs) are used to indicate the occurrence of LAMP. Its products are mixed with AuNPs in an optimized ration, upon which the deoxyribonucleotides (dNTPs) bind to the AuNPs via ligand-metal interactions and thus enhance their stability. If a target DNA is amplified, the vast reduction of the dNTPs leads to the aggregation of AuNPs and a color change from red to blue. The success of the method strongly depends on the ionic strength of the solution and the initial concentration of dNTPs. Unlike other methods for identification of isothermal products, this method is simple and can be readily applied at sites where instrumentation is inadequate or even lacking. Key words: label-free; gold nanoparticle; loop-mediated isothermal amplification; naked eyes. 0 Introduction Gold nanoparticles based colorimetric assays have shown great promise for visualization of genetic variation [1, 2] its size-dependent and distance-dependent optical properties [3, 4] were cross-linked to target DNA [5] or at a high ionic strength [6] 25 30 aggregated AuNPs broadens and red-shifts. The large color shift can be easily read out with naked eyes as an end point detection method; therefore, no expensive instruments are required. With these characteristics, AuNPs based colorimetric strategies were suitable to create a fast, specific and low-cost analysis platform for point of care diagnostics [7]. To detect the target DNA at low levels, an amplification step is usually required. The combination of AuNPs based assays with conventional PCR technique has brought significant improvement compared with conventional PCR technologies [8, 9] cycling device restr