Chen Wei, Jiang Yuan, Ji Baoqing, Zhu Changqin, Liu Liqiang, Peng Chifang, Jin M Kim, Qiao Ruirui, Jin Zhengyu, Wang Libing, Zhu Shuifang, Xu Chuanlai
School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, PR China.
Biosens Bioelectron. 2009 May 15;24(9):2858-63. doi: 10.1016/j.bios.2009.02.015. Epub 2009 Feb 27.
An ultrasensitive and rapid sequential injection analysis (SIA) based on real-time PCR (SIA-rt-PCR) assay was developed by using different nanoparticles for the detection of small molecule chemicals residues. Gold (Au) nanoparticle, conjugated with goat anti-rabbit IgG and duplex strand DNA (dsDNA), was used as a substitute for chemiluminescent probes in an SIA system. By indirect competitive immunoreactions in the SIA system, the gold nanoparticles were attached to antigens which were immobilized by superparamagnetic nanoparticles (SMNPs). The dsDNA on the gold nanoparticles was dehybridized and then the single-stranded DNA (ssDNA) was collected and quantified with rt-PCR, which showed a rather low linearity range from 2.5 attomol L(-1) (aM) to 250 femtomol L(-1) (fM) and the LOD was 1.4 aM. This method, which is rapid, automated and capable of high-throughput, was used to detect methyl-3-quinoxaline-2-carboxylic acid (MQCA) residues in real samples. The analytical results had a coefficient of variation of less than 15% and the recovery was 89-108%.
通过使用不同的纳米颗粒来检测小分子化学物质残留，开发了一种基于实时聚合酶链反应（SIA-rt-PCR）的超灵敏快速顺序注射分析（SIA）方法。与山羊抗兔免疫球蛋白G和双链DNA（dsDNA）偶联的金（Au）纳米颗粒，被用作SIA系统中化学发光探针的替代品。通过SIA系统中的间接竞争免疫反应，金纳米颗粒附着到由超顺磁性纳米颗粒（SMNP）固定的抗原上。金纳米颗粒上的dsDNA被解链，然后收集单链DNA（ssDNA）并用rt-PCR进行定量，其线性范围相当低，为2.5阿托摩尔每升（aM）至250飞摩尔每升（fM），检测限为1.4 aM。该方法快速、自动化且能够高通量，用于检测实际样品中的3-甲基喹喔啉-2-羧酸（MQCA）残留。分析结果的变异系数小于15%，回收率为89%-108%。
