三种纳米金颗粒对聚合酶链式反应扩增效率和特异性的影响

比较了采用柠檬酸三钠(Trisodium citrate,Na3Ct)还原法、聚乙烯吡咯烷酮(Polyvinylpyrrolidone,PVP)包裹法和巯基丙酸-同型半胱氨酸(Mercaptopropionic Acid-Homocystine,MPA-HCys)包裹法制备的3种纳米金颗粒(Gold nanoparticles,AuNPs)对聚合酶链式反应(Polymerase Chain Reaction,PCR)扩增效率及特异性的影响,以评估金纳米材料表面性质与DNA生物大分子之间的相互作用.实验结果显示,3种AuNPs均能显著提高PCR效率,它们提高PCR效率的能力依次为:Na3 Ct-AuNPs(12 nm)MPA-HCys-AuNPs(12 nm)PVP-AuNPs(4 nm),而且这种增强效应与其浓度和尺寸相关.相同实验条件下,具有相似尺寸大小的Na3Ct-AuNPs比MPA-HCys-AuNPs更能有效提高PCR效率,这说明AuNPs的表面性质也是影响PCR效率的重要因素之一.另外,3种AuNPs均能有效消除竞争性引物对PCR的抑制作用,并且这种作用与其浓度密切相关.上述研究结果表明,具有不同表面性质的3种AuNPs均能有效提高PCR效率和特异性,并且这种作用与其浓度、尺寸和表面性质密切相关.

Effect of gold nanoparticles on the efficiency and specificity of polymerase chain reaction

In this report,three types of gold nanoparticles(AuNPs) were prepared by the method of sodium citrate(Na3Ct) reduction,polyvinylpyrrolidone(PVP) packaging and mercaptopropionic acid(MPA),homocystine(HCys) surface modification to compare their effect on the Polymerase Chain Reaction(PCR),and to evaluate the interaction between the surface of the AuNPs and DNA molecules.The results showed that the three types of AuNPs significantly enhanced PCR efficiency in the order of Na3Ct-AuNPs(12 nm)>MPA-HCys-AuNPs(12 nm)>PVP-AuNPs(4 nm),and the enhancement was dependent on their concentration and size.Furthermore,by comparing two nanoparticals of similar size,Na3Ct-AuNPs and MPA-HCys-AuNPs,we found Na3Ct-AuNPs improved the PCR efficiency more profoundly,indicating that the surface property was also one of the important factors in influencing the PCR efficiency.In addition,the three AuNPs effectively eliminated the inhibition of competitive primer on PCR in a concentration-dependent manner.These results indicated the three AuNPs could dramatically enhance the efficiency and specificity of PCR,and this effect was related to its concentration,size and surface property.