应用不对称PCR法进行表面等离子共振微生物核酸检测

Detection of Surface Plasmon Resonance on Microorganism's DNA with Asymmetric PCR Method

目的通过比较不同方法获取的微生物核酸样本用于表面等离子共振(SPR)传感器检测效果的差异,对不对称PCR法制备核酸单链用于SPR检测效果进行研究。方法分别将经淬火处理和未经处理的不对称PCR、常规PCR产物用于SPR传感器检测,以检测曲线和检测响应为指标对各样本在SPR传感器上的检测效果进行比较。结果实验结果表明不对称PCR可以直接获得核酸单链,较常规PCR方法在SPR核酸检测上具有更好的检测效果:经过淬火处理后不对称PCR产物样本平均折射率(R I)改变为39.33(RU),而相同浓度的普通PCR产物只有10.63(RU);同时在SPR实时检测曲线也可以观察到不对称PCR产物能够产生明显的结合信号,而普通PCR则没有类似信号出现。结论利用不对称PCR方法可以制备直接用于SPR检测的核酸单链,并且在杂交检测效果上明显优于普通PCR方法。

Objective To study the effects of SPR detection on single-strand DNA with asymmetric PCR meth- od, by comparing the differences of detection effects of surface plasmon resonance （SPR） sensors on nucleic acid samples obtained from microorganisms with different PCR methods. Methods PCR products obtained from asymmetric or symmetric PCR were treated with quenching or not respectively and then used in SPR detection. The detection effects were compared with detection curves and responses of refractive index（RI）. Results The results showed that single-strand DNA could be obtained directly from asymmetric PCR. SPR sensor curves ob- tained from asymmetric PCR products were better than that from symmetric PCR products. The average shift of RI of asymmetric PCR product was 39.33 （RU）, while △RI of symmetric PCR product was only 10.63 （RU）. High hybridization signal could be found in was observed in symmetric PCR product detection asymmetric PCR can be used for SPR detection and asymmetric PCR product detection and no obvious signal Conclusion Single-strand DNA directly obtained from get better effects than that from symmetric PCR method.