应用短串联重复序列快速诊断唐氏综合征  

Rapid diagnosis of Down syndrome use short tandem repeats

作　　者：何薇[1] 林琳华[2] 吴菁[1] 尹爱华[1] 姚秋璇[2] 

机构地区：[1]广东省妇幼保健院广州医学院附属广东省妇儿医院产前诊断中心,广州510010 [2]深圳市人民医院产前诊断中心,深圳518020

出　　处：《中国优生与遗传杂志》2013年第10期36-39,共4页Chinese Journal of Birth Health & Heredity

摘　　要：目的在实时荧光定量PCR技术的基础上建立一种诊断效果较好、简便、经济的快速诊断唐氏综合征的技术。方法以D21s1411,D21s1259为检测位点,D19s222为参照位点,用SYBR GreenⅠ作为荧光染料,采用荧光定量PCR技术检测37例唐氏综合征及131例非唐氏综合征胎儿羊水标本的STR片段,分析唐氏综合征组及非唐氏综合征组的4对△Ct值。结果 1.非唐氏征组的△Ct1411及△Ct1259的均数、标准差及95%置信区间均为正值,唐氏征组的△Ct1411及△Ct1259的均数及95%置信区间均为负值;两组间95%置信区间互不重叠。2.非唐氏征组与唐氏征组位点D21s1411及D21s1259△Ct值差异均有统计学意义,分别为t=12.088,P=0.000及t=13.884,P=0.000。3.用D21s1411进行诊断时,ROC曲线下的面积为0.9291,将△Ct14110.1327为截止值时的灵敏度与特异度之和最大;用D21s1259进行诊断时,ROC曲线下的面积为0.985,将△Ct1259=-0.2016作为截止值时的灵敏度与特异度之和最大。结论 1.以D21s1411和D21s1259为检测位点,应用荧光定量PCR方法检测羊水标本的STR片段,可有效判断胎儿是否为唐氏综合征,检出率达91%以上。2.用SYBRGreenⅠ作为染料进行荧光定量PCR检测,操作步骤简单,成本较低,效果良好。Objective： To establish a short tandem repeats （STR） sequence detection technique using the real -time fluorescence quantitative polymerase chain reaction （PCR）, and this technology has a good diagnosis effect, convenient and economy. Methods： The amniotic fluid samples were collected from 37 fetus with DS and 13l non - DS fetus who were diagnosed using the standard cytogenetic method. We detected three STR sequences by real -time fluorescence quantitative PCR, using D21s14l 1 and D21 s1259 on chromosome 21 as detection loci, D19s222 on chromosome 19 as the control locus, and SYBR Green I as fluorescent dye. Four pairs of Ct values were analysed between the DS group and the non - DS group. Results： 1. The mean value and the 95% confidence interval of △Ct1411 and △Ct1259 were positive in non- DS group, while they were negative in DS group. The 95% confidence intervals did not overlap between the two groups. 2. The difference of △Ct1411 between the DS group and non - DS group was greatly significant （ t1411 = 12. 088, P1411 =0. 000）. The difference of △Ct1259 between two groups was significant too （t1259 = 13. 884, P,259 =0. 000）. 3. The area under the ROC curve was 0. 929 when using D21s1411 as STR marker. When the cut - off value of △Ct1411 was 0. 1327, the sensitivity add the specificity is maximum. While the area under the ROC curve was 0. 985 when using D21s1259 as STR marker. When the cut - off value of △Ct1259 was - 0. 2016, the sensitivity add the specificity is maximum. Conclusions 1. The detection of the two STR sequences （D21s1411 and D21s1259） on chromosome 21 from amniotic fluid samples by real -time fluorescence quantitative PCR technique can effectively diagnose DS. The detection rate was greater than 91%. 2. The detection technique by real - time fluo- rescence quantitative PCR using SYBR~ Green I as fluorescent dye was simple, economical and reliable.

关 键 词：短串联重复序列 荧光定量PCR 唐氏综合征 产前诊断 SYBR Green I 

分 类 号：R714.55[医药卫生—妇产科学]