摘要
目的分析1例D—变异体的分子背景。方法分别采用血清学方法和序列特异性引物PCR(PCR-SSP)方法检测标本RhD、C、c、E、e抗原和RHCE基因,同时序列分析RHAG、RHCE基因全长编码区序列,最后进行RHD合子型分析和家系分析。结果先证者血清学测定Rh表型为D—,PCR-SSP结果为DC-;RHAG第1-10外显子测序显示,编码区与Gen Bank NC-000006标准序列一致,未观察到形成Oldeide和Duclos-Like抗原的碱基变异,亦未见形成高频抗原Duclos(RHAG1)的RHAG316C>G突变,所有序列均未见杂合峰;RHCE基因第1-10外显子扩增显示,缺失第3-5外显子,其余外显子序列与标准序列一致,分析第1、2外显子序列为RHC,提示该等位基因为RHCE(e)-D(3-5)-CE(e);家系分析其父母、弟弟的RHAG序列与先证者完全一致,父母Rh血清学表型分别为DCCee和Dcc EE,其父亲的PCR-SSP和测序结果一致,其母亲PCR-SSP结果 DCc EE与测序结果一致,而与血清学结果不符(Dcc EE),其弟血清学表型、PCR-SSP和测序结果,均与先证者一致,结合RHD合子型分析结果,该家系父母分别为DCe/DC-和Dc E/DC-基因型,先证者和弟弟为DC-/DC-。结论 RHCE-D(3-5)-CE等位基因形成D—表型并稳定遗传。
Objective To study the genetic background of a D--variant. Methods The D, C, c, E and e antigens were tested by the routine serological methods. A sequence-specific primer PCR （PCR-SSP） method was applied for detection of RHCE genotype. The whole length coding region of RHAG and RHCE was sequenced respectively. Furthermore, the RHD zygosity was also detected with a PCR method. Results The proband sample was determined as D--serologically, but DC-by PCR-SSP. The RHAG coding region sequencing results showed that 10 exons＇ sequences were consistent with RHAG standard sequence （ GenBank NC-000006）. There were no mutations observed for Oldeide and Duclos-Like antigens, nor the mutation of 316C 〉 G found in high frequency antigen Duclos. The RHCE sequencing results showed that exons 3, 4 and 5 were deleted, and the other sequences were identical with a common RttCE sequence. Since the exons 1 and 2 were RHC sequences, a RHCE （e） -D （3-5） -CE （e） allele was identified. Family investigation showed that the RHAG sequences of parents and brother were identical to the proband＇s. The Rh phenotypes of parents were DCCee and DccEE respectively, which was consistent with the PCR-SSP and sequencing results of the father＇s, but the mother＇s was DCcEE in genetic test. The brother＇s was consistent with the proband completely. Combining the zygosity results, the genotypes of parents were DCe/DC- and DcE/DC-, and both the proband＇s and the brother＇s were DC-/DC-. Conclusion The RHCE-D（3-5）-CE allele produces the D--variant and inherits stably in the investigated family.
作者
陈文伟
刘圆圆
唐朝晖
易品
黎诚耀
张印则
徐华
邵超鹏
CHEN Wenwei LIU Yuanyuan TANG Zhaohui Ylpin LI Chengyao ZHANG Yinze XU Hua SHAO Chaopeng(School of Biotechnology , Southern Medical University, Guangzhou 510515 ,China Gansu Blood Center Shenzhen Blood Center Shaanxi Blood Center The First Affiliated Hospital of Shenzhen University.)
出处
《中国输血杂志》
CAS
北大核心
2016年第8期788-790,共3页
Chinese Journal of Blood Transfusion
基金
深圳市科技创新计划(JCYJ2014041470821221
JCYJ20130401110246572)
