等位基因特异性实时荧光PCR技术进行α0-地中海贫血的排除性无创产.DOC

等位基因特异性实时荧光PCR技术α0-地中海贫血的无创产前诊断 刘彦慧1,吴亚敏2, 石少权3,娄季武1, 马秋林1, 何怡1, 徐婉芳1, 林洋洋1 ,周万军4* 523700东莞东莞市妇幼保健院产前诊断中心1,523220广东东莞东华医院中心实验室2,519000广东珠海中山大学第五附属医院妇产科3,510515广东广州南方医科大学基础医学院遗传学教研室4 * 通信作者: Email: zhouwanjun72@163.com 【摘要目的 建立纯合α0-地中海贫血无创产前诊断等位基因特异性实时荧光PCR技术AS-qPCR)。方法PCR微测序技术高风险缺失区内9个SNP差异位点AS-qPCR技术检测差异结果 167个家系160个家系找到1个或多个有的SNP位点，检测率约为95.8%94例检测父源性正常等位基因57.6%；所有家系的绒毛或羊水基因检测结果与本方法的符合率为100%。 结论AS-qPCR检测孕妇血浆DNA中双亲差异位点，在孕早期即可进行纯合α0-地中海贫血的无创产前诊断，可使约50%高孕产妇免创伤性诊断。 关键词α-地中海贫血；无创产前诊断；游离；缺失 基金项目东莞医疗卫生单位重点科研课题资助项目(200910515024) Non-invasive exclusion prenatal for homozygous (0-thalassemia by allele-specific real-time fluorescence PCR Liu Yanhui1, 　Wu Yamin2,　Shi Shaoquan3,　Lou Jiwu1，Ma Qiulin1,He Yi1,　Xu Wanfang1,　Lin Yangyang1,　Zhou Wanjun4*　（1 prenatal diagnosis center, Maternal and Child Health Hospital of DongGuan, GuangDong, 523700,china. 2 Center Laboratory, Donghua Hospital, Dongguan,GuangDong, 523220,China. 3 The Fifth Affiliated Hospital, Sun Yat-Sen University, ZhuHai, GuangDong, 519000, China. 4 Department of Genetics，Southern Medical UniversityGuangZhou, GuangDong, 510515, China.） 【Abstract】 Objective To establish and evaluate an allele-specific real-time fluorescence PCR (AS-QPCR) assay for homozygous a0-thalassemia exclusive non-invasive prenatal diagnosis. Methods Among 9 SNPs within the SEA deletion range, the different SNPs between parents were determined with PCR micro-sequencing technology. The paternal SNP was detect in maternal plasma DNA with AS-qPCR method to determine whether the paternal normal haplotype inherited to the fetus. Results 160 of 167 families were found have one or more parents-difference SNPs with the detection rates over 95.8%. The paternal normal SNPs were detected in 94 maternal plasma DNA and then they avoid invasive prenatal diagnosis with the detect rate of 57.6%. This results were 100% coincidence with the results of CVS or amniocentesis. Conclusion Using AS-QPCR method detect the parents-difference SNPs in maternal plasma DNA