摘要
目的通过对两例散发的BMD家系中的先证者和胎儿进行DMD基因和SRY基因分析,拟建立一种快速、准确、适用于临床的DMD/BMD的产前基因诊断方法。方法提取羊水细胞基因组DNA,通过检测SRY基因鉴定胎儿性别,以多重PCR方法对DMD基因缺失热区的18个外显子进行基因缺失的筛查,结合4对STR引物(内含子44、45、49、50)进行短串联重复序列多态性连锁分析(STR-PCR),分析结果经DNA测序验证。结果 SRY基因分析结果显示家系1胎儿为女性,家系2胎儿为男性,与超声诊断结果一致。两例BMD先证者均未检测到18个外显子的缺失,两家系的母亲及家系1的女性胎儿均为BMD基因STR-49位点杂合子,家系2的男性胎儿为BMD。同时发现了STR45、STR50的非常见基因型。结论应用SRY基因检测胎儿性别及多重PCR方法对DMD基因外显子缺失的检测,结合STR-PCR技术检出非缺失型携带者并行产前诊断,此三联PCR方法具有快速、简便、直观可靠的特点,特异性高,实验可操作性强,适用于临床推广应用。
Objective In order to establish a fast,accurate and suitable DMD/BMD clinical genetic diagnosis before antepartum,we execute DMD gene and the SRY gene analysis through proband and fetus of two sporadic BMD family.Methods Amniotic fluid cells genomic DNA was extracted;fetal gender could be identified by SRY gene testing;screening of gene deletion of the DMD gene deletion in hot area of 18 exons by multiplex PCR method;in addition to four pairs of STR primers(introns 44,45,49,50),short tandem repeat polymorphism linkage(STR-PCR)was be analysis,the final analysis results was verified through DNA sequencing testing.Results SRY gene analysis showed the fetus of family 1 was a female and that of family 2 was a male,the results were consistent with ultrasound diagnosis.Two patients with BMD proband was not detected in the deletion of exon 18,1 female fetus of the mother and family of the two families were the BMD gene STR-49 points heterozygous male fetus of family 2 for the BMD.Meanwhile,the uncommon genetype of STR45,STR50 was found also.Conclusions It is demonstrated that the application SRY gene detection of fetal gender and multiplex PCR method for the DMD gene deletion of exon detection,combined with the STR-PCR technology to the detection of non-deletion carriers parallel prenatal diagnosis,this triple PCR method is fast,simple and intuitive reliable characteristics of the high specificity,the experiment may be feasible,and applicable to clinical application.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第21期6766-6771,共6页
Chinese Journal of Clinicians(Electronic Edition)
基金
武汉市科技攻关项目(200760423158)
