Automated and ultrasensitive detection of methyl-3-quinoxaline-2-carboxylic acid by using gold nanoparticles probes SIA-rt-PCR.
Automated and ultrasensitive detection of methyl-3-quinoxaline-2-carboxylic acid by using gold nanoparticles probes SIA-rt-PCR.
复制标题
使用金纳米粒子探针 SIA-rt-PCR 对甲基-3-喹喔啉-2-羧酸进行自动超灵敏检测。
DOI：
10.1016/j.bios.2009.02.015
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发表时间：
2009-05
期刊：
影响因子：
--
通讯作者：
Jin Zhengyu
中科院分区：
文献类型：
--
作者：
Chen Wei;Wang Libing;Zhu Shuifang;Xu Chuanlai;Jiang Yuan;Ji Baoqing;Zhu Changqing;Liu Liqiang;Peng Chifang;M. Kim Jin;Qiao Ruirui;Jin Zhengyu
建立了一种基于实时荧光定量PCR（SIA-rt-PCR）的超灵敏、快速的顺序注射分析（SIA）方法，用于小分子化学品残留的检测。将羊抗兔IgG和双链DNA（dsDNA）偶联的金（Au）纳米粒子作为荧光探针用于SIA体系。通过SIA系统中的间接竞争免疫反应，金纳米颗粒被附着到由超顺磁性纳米颗粒（SMNP）固定的抗原上。将金纳米颗粒上的dsDNA去杂交，然后收集单链DNA（ssDNA），并用rt-PCR进行定量，其显示出相当低的线性范围为2.5attomolL−1（aM）至250 femtomolL−1（fM），LOD为1.4aM。该方法具有快速、自动化程度高、高通量等优点，可用于真实的样品中甲基-3-喹喔啉-2-羧酸（MQCA）残留的检测。分析结果的变异系数小于15%，回收率为89- 108%。
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 2.5attomolL−1(aM) to 250 femtomolL−1(fM) and the LOD was 1.4aM. 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%.
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