双重相对定量SD-qPCR在快速诊断性染色体变异中的应用  

Rapid determination of sex chromosome abnormality using double relative quantitative SD-qPCR

作　　者：孙雷 龙驹 樊祖茜 吴素萍 林桂先 凌秀明 SUN Lei;LONG Ju;FAN Zuqian;WU Suping;LIN Guixian;LING Xiuming(Xi'an Jiaotong University Health Science Center,Xi'an 710061,Shaanxi,China;Laboratory of Medical Genetics,Qinzhou Maternal and Child Health Hospital,Qinzhou 535099,Guangxi,China;Department of Clinical Laboratory,Qinzhou Maternal and Child Health Hospital,Qinzhou 535099,Guangxi,China)

机构地区：[1]西安交通大学医学部,陕西西安710061 [2]钦州市妇幼保健院基因与遗传实验室,广西钦州535099 [3]钦州市妇幼保健院检验科,广西钦州535099

出　　处：《检验医学》2022年第6期557-560,共4页Laboratory Medicine

基　　金：国家自然科学基金项目(81660259);广西壮族自治区卫生健康委员会自筹经费科研课题(20221295)。

摘　　要：目的建立一种可进行双重相对定量的基于重复序列的荧光定量聚合酶链反应(SD-qPCR)方法,实现单管同时检测X和Y染色体数目。方法建立双重相对定量SD-qPCR检测体系,并将其用于197例羊水样本的检测。根据重复序列间的相对定量2-ΔΔCt值来判断样本X和Y染色体的数目,将检测结果与染色体核型分析结果进行比较。结果筛选出了2对重复序列分子标记,分别定位于16号染色体、X染色体和1号染色体、Y染色体;197例羊水样本中,性染色体数目正常样本185例;45,X样本5例;47,XXX样本3例;47,XXY样本2例;47,XYY样本2例。双重相对定量SD-qPCR检测结果与染色体核型分析结果一致。结论建立的双重相对定量SD-qPCR方法可实现单管同时检测X和Y染色体的数目,具有检测位点多、检测结果稳定的特点,可在临床推广使用。Objective To establish a single tube fluorescent double relative segmental duplication-quantitative polymerase chain reaction(SD-qPCR)for determining the number of chromosome X and Y simultaneously.Methods The established SD-qPCR was used for the determination of 197 amniotic fluid samples.The numbers of chromosome X and Y were determined by the relative quantitative 2-ΔΔCt value between segmental duplications.The results are compared with chromosome karyotype analysis.Results Two pairs of segmental duplication molecular markers were screened and located on chromosome 16 and chromosome X,chromosome 1 and chromosome Y,respectively.Among the 197 samples,there were 185 samples with normal sex chromosome,5 cases of 45,X,3 cases of 47,XXX,2 cases of 47,XXY and 2 cases of 47,XYY.The results of SD-qPCR were consistent with those of chromosome karyotype analysis.Conclusions The established SD-qPCR can determine the numbers of chromosome X and chromosome Y in a single tube simultaneously.It has the advantages of multiple loci and stability,and it can be widely used in clinic.

关 键 词：产前诊断 非整倍体 实时荧光定量聚合酶链反应 性染色体 

分 类 号：R446.1[医药卫生—诊断学]