两种不同检测方案在α-地中海贫血植入前遗传学诊断中的应用分析  

Application of two different detection schemes in preimplantation genetic diagnosis of alpha-thalassemia

作　　者：严提珍 李伍高 李哲涛 唐永梅 蔡稔 韦立红 秦祖兴 范莉 李忻琳 YAN Ti-zhen;LI Wu-gao;LI Zhe-tao;TANG Yong-mei;CAI Ren;WEI Li-hong;QIN Zu-xing;FAN Li;LI Xinlin(Department of Medical Genetics,Liuzhou Municipal and Child Healthcare Hospital,Liuzhou Key Laboratory of Birth Defect Prevention and Control,Guangxi,Liuzhou,545001;Reproductive Centre of Liuzhou Municipal and Child Healthcare hospital,Liuzhou Marternal and Children Healthcare Hospital,Guangxi Guangxi,Liuzhou,545001)

机构地区：[1]柳州市妇幼保健院医学遗传科,柳州市出生缺陷预防与控制重点实验室,广西柳州545001 [2]柳州市妇幼保健院生殖健康助孕中心,广西柳州545001

出　　处：《中国优生与遗传杂志》2020年第2期233-235,共3页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学资金(81360091);广西壮族自治区卫生和计划生育委员会自筹经费科研课题(Z20170527);柳州市科学研究与技术开发计划课题(2014G020404、2017BH20312和2018AF10501)。

摘　　要：目的比较两种不同的检测方案在α-地中海贫血胚胎植入前遗传学诊断(preimplantation genetic diagnosis,PGD)中的应用。方法对2017年1月至2018年12月在本院进行α-地中海贫血PGD的囊胚检测结果进行回顾性分析,其中蛋白酶K裂解后荧光PCR直接检测致病位点方法(方案1)40个周期,全基因组扩增结合短串联重复序列单体型分析和致病位点检测方案(方案2)108个周期,将两组的诊断结果与随访结果进行比较。结果方案1检测成功率为95.15%(16/330),正常胚胎为21.97%(69/314),携带者胚胎为50.64%(159/314),重型地贫胚胎为27.39%(86/314);方案2检测成功率为93.02%(54/774),正常胚胎为19.17%(138/720),携带者胚胎为44.72%(322/720),重型地贫胚胎为25.83%(186/720),其他类型(包括疑似单体10个,疑似三体15个,疑似重组49个)胚胎为10.28%(74/720)。两种方案的检测成功率、正常胚胎率、携带者胚胎率、重型地贫胚胎率均无显著性差异(P>0.05)。但方案1不能检出16号染色体拷贝数异常及HBA基因附近重组情况。结论全基因组扩增结合短串联重复序列单体型分析和致病位点检测方案较蛋白酶K裂解后荧光PCR直接检测致病位点方案更适合进行α-地中海贫血PGD。Objective:To compare the application of two different detection schemes in preimplantation genetic diagnosis(PGD)of alpha-thalassemia embryos.Methods:The blastocyst detection results of alpha-thalassemia PGD in our hospital from January 2017 to December 2018 were retrospectively analyzed.Among them,40 cycles were used for direct detection of pathogenic sites by fluorescent PCR after protease K cleavage(scheme 1),108 cycles were used for genome amplification combined with short tandem repeat sequence haplotype analysis and pathogenic sites detection(scheme 2).The follow-up results were compared.Results:The success rate of scheme 1 was 95.15%(16/330).21.97%(69/314)embryos were normal,50.64%(159/314)embryos were carrier,and 27.39%(86/314)embryos were severe thalassemia.The success rate of scheme 2 was 93.02%(54/774).19.17%(138/720)embryos were normal,44.72%(322/720)embryos were carrier,25.83%(186/720)embryos were severe thalassemia.Other types of embryos(including 10 suspected monomers,15 suspected trisomies and 49 suspected recombinants)were 10.28%(74/720).There was no significant difference in the success rate,normal embryo rate,heterozygosity embryo rate and severe thalassemia embryo rate between the two schemes(P>0.05).However,scheme 1 could not detect abnormal copy number of chromosome 16 and recombination near HBA gene.Conclusion:Genome-wide amplification combined with short tandem repeat sequence haplotype analysis and pathogenic site detection is more suitable for alpha-thalassemia PGD than direct detection of pathogenic site after protease K cleavage by fluorescent PCR.

关 键 词：Α-地中海贫血 植入前遗传学诊断 短串联重复 单体型分析 全基因组扩增 

分 类 号：R726.2[医药卫生—儿科]