单细胞巢式PCR检测RHD的初步研究  

Primary study on single-cell nested PCR for detection of RHD

作　　者：刘建强[1] 吴大洲[1] 张印则[2] 周华友[3] 徐华[1] 

机构地区：[1]陕西省血液中心,陕西西安710061 [2]深圳市血液中心 [3]广州中医药大学第二附属医院

出　　处：《中国输血杂志》2015年第7期760-762,共3页Chinese Journal of Blood Transfusion

摘　　要：目的建立单细胞巢式PCR检测技术,用以检测单个细胞的RHD。方法采用稀释法获得单个细胞,直接提取DNA,根据RHD特点设计两对特异性引物进行巢式PCR,凝胶电泳观察结果,将扩增产物测序以检测扩增的目的基因。结果通过巢式PCR,阳性对照可以获得明显的目的产物,阴性和空白对照无扩增产物,10份Rh阳性单细胞检测标本均能检出RHD。结论建立的检测RHD单细胞巢式PCR检测技术具有很好的特异性和敏感性,可应用于生殖医学等领域,为预防Rh新生儿溶血病提供新的技术途径。Objective To establish a single-cell nested PCR method to detect RHD. Methods Individual cells were obtained by dilution method,and DNA was isolated directly. Nested PCR was conducted by two pairs of primers which were designed according to RHD specificity. The results of gel electrophoresis were observed,and the amplified products were sequenced. Results By nested PCR,the positive control could obtain obvious target products,while the negative and the blank control did not amplify product. RHD were detected from 10 samples of Rh positive single cell. Conclusion The technology of single-cell nested PCR to detect RHD of single cell was established with good specificity and sensitivity,which could be applied in the field of reproductive medicine to provide a new technical way to prevent Rh HDN.

关 键 词：单细胞巢式PCR RHD 生殖医学 RH新生儿溶血病 

分 类 号：R457.11[医药卫生—治疗学]