SPR技术在产前检测胎儿RhD血型基因中的应用

The application of SPR technology in prenatal testing of fetal RHD blood type gene

目的探讨利用表面等离子体共振技术(SPR)检测胎儿RhD血型基因的可行性,建立一种胎儿RhD血型基因快速诊断的新方法。方法采用氨基偶联法在SPR芯片表面固定不同种类DNA对应的RNA探针,并优化芯片分析条件,再运用RNase H酶水解,放大检测信号,确定该方法检测的条件;再用RhD血型基因第5,7外显子的RNA探针检测其相应的DNA分子,分析SPR芯片检测信号的特异性与灵敏性。结果SPR技术检测RhD血型基因第5,7外显子的灵敏度与特异性均较好,RhD血型的RNA探针在SPR芯片上可特异性地检测RhD血型基因第5,7外显子,SPR信号检测RhD血型基因DNA分子的浓度为100pmol/L。结论SPR技术可快速检测RhD血型基因的相应外显子,且SPR技术操作简便快速、可靠直观,且非标记,可为RhD阴性孕妇产前预测胎儿RhD血型基因提供一种新方法。

Objective To study the feasibility of detecting fetus RhD type gene by Surface Plasmon Resonance(SPR)technology,and to establish a new rapid diagnosis method for fetus RhD type gene.Methods The different types of DNA corresponding RNA probes were fixed on the surface of SPR chip by using amino coupling methods,and optimize the chip analysis condition,and then using the RNase H enzyme hydrolysis,signal amplification detection,at last the detection conditions were determined.We use the RhD type gene exon 5,7 of RNA probe to test its corresponding DNA molecules,and analyse the specificity and sensitivity of SPR chip detection signal.Results The SPR technique for detecting the exon 5,7 of RhD blood type gene shows good sensitivity and specificity in all,SPR technology can specifically detect the Exon 5,7 of RhD blood type gene,and the sensitivity of for detecting RhD gene exon 5,7 is 100 pmol/L by SPR.Conclusion The SPR technology can quickly detect RhD gene accordingly,SPR technology is simple,rapid,reliable and label-free,which can provide a new way predicting fetal RhD type for RhD negative prenatal.