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根据猪流行性乙型脑炎病毒(JEV)E基因的保守区域设计1对扩增270bp片段的特异性检测引物,将纳米金颗粒添加到PCR反应体系中,同时优化反应条件,建立了高效、灵敏的纳米PCR检测方法。采用该方法灵敏度可达3.26×10~2拷贝/μL,普通PCR方法仅为3.26×10~4拷贝/μL,且与其他病毒没有交叉感染。试验结果表明,该纳米PCR检测方法不仅特异性好,而且其敏感性是普通PCR检测方法的100倍。采用纳米PCR方法和普通PCR方法对临床送检的32份样品进行检测,检测结果纳米PCR检出2份阳性样品,普通PCR检测均为阴性。
According to the conserved region of porcine encephalitis virus (JEV) E gene, a pair of specific primers was designed to amplify a 270bp fragment. The gold nanoparticles were added to the PCR reaction system, and the reaction conditions were optimized. An efficient, Sensitive nano-PCR detection method. The sensitivity of this method can reach 3.26 × 10 ~ 2 copies / μL, the average PCR method is only 3.26 × 10 ~ 4 copies / μL, and no cross-infection with other viruses. The experimental results show that the PCR method is not only specific, but also sensitive to 100 times PCR. Nano-PCR and PCR were used to detect 32 clinical samples. The results of PCR showed that two positive samples were detected by Nano-PCR. All the PCR tests were negative.