应用纳米金探针检测HSV-2的目视化基因芯片

本实验建立了一种应用金标链霉亲和素探针的目视化高灵敏度检测单纯疱疹病毒2型(HerpesSimplexVirus-2,HSV-2)的基因芯片。该芯片以HSV-2DNA聚合酶的高保守区为靶序列,设计HSV-2特异性引物和探针,通过PCR反应使扩增产物标记上生物素;氨基修饰的探针固定在活化的玻片上,与生物素标记的扩增产物杂交;利用生物素与链酶亲和素高亲合力的特性,加入纳米金标记的链酶亲和素后形成生物素-链酶亲和素-纳米金生物反应放大系统;银染反应后,达到目视化检测HSV-2效果。该HSV-2检测基因芯片能目视化检测出100fmol/L的HSV-2扩增产物,具有灵敏度高,低成本的特点,通过临床标本验证表明该芯片具有较高的准确性。

A Visual Gene Array for the Detection of HSV-2 Based on Nanogold Probes

A visual HSV-2 detection gene array was developed based on nanogold probes. The specific primer and probe of HSV-2 were designed based on the highly conservative region of HSV-2 DNA polymerase gene and the HSV-2 PCR amplicon were labeled by biotin through PCR reaction. Amino-modified probes were spotted on an activated glass surface on which hybridized with the PCR amplicon. Subsequently, the streptavidin-nanogold was added to form the amplification system of the biotin-streptavidin-nanogold complex. At last, the step of silver staining amplified the signal of hybridization, Therefore it was easy to detect the HSV-2 by naked eyes. As little as 100fmol/L of HSV-2 amplicon could be detected on the array, The HSV-2 detection gene array can detect the HSV-2 virus infection with high sensitivity and low cost.