对孕妇外周血中的单个有核红细胞及游离DNA来源的鉴定

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1、•论著・对孕妇外周血屮的单个有核红细胞及游离DNA来源的鉴定陈汉平王陶然贺桂芳卢运萍马庭元【摘要】目的探讨孕妇外周血屮单个胎儿有核红细胞及游离DNA在非创伤性产前诊断的可行性。方法对116例孕妇外周血进行检测。（1）对51例14〜26孕周的妇女外周血经密度梯度离心后用显微操作分离单个有核红细胞。⑵捉収65例孕妇（5〜40孕周）外周血血浆DNAo（3）应用巢式聚合解链反应（PCR）扩增单个右核红细胞的男性SRY基因，应用引物延伸预扩増（PEP）法及巢式PCR及游离的DNA可來自胎儿，它可成为产前诊断的胎儿物质來源。（2）单细胞分离技术使孕妇外周血中的胎儿有核红

2、细胞的分选纯度几乎达到100%，解决了母胎细胞混合的难题，为非创伤性产前诊断提供了一条新思路。【关键词】胎儿；红细胞；性别预选；聚合酶链反应Identificationoftheoriginofsinglenucleatedredbl(H)dcellsandfreeDNAinperipheralbloodofpregnantwomenCHENHanping,WANGTaoran,HEGuifang,LUYunping,MATingyuan.DepartmentofGynecologyandObstetrics.TongjiHospital,TongjiMed

3、icalCollege.HuazhongUniversityofScienceandTechnology,Wuhan430030China[Abstract]ObjectiveToinvestigatethefeasibilityofusingsinglefetalnucleatedredbloodcells(FNRBCs)andfreeDNAfornoninvasiveprenataldiagnosis.Methods116samplesofmaternalbloodwereanalyzed.SingleFNRBCswereisolatedfromthep

4、eripheralbloodsamplesof51pregnantwomenwiththegestationalperiodof14to26weeksbymicromanipulationtechniquesafterdensitygradientcentrifugation.NestedpolymerasechainreactionmethodwasusedtoamplifytheSRYgene・PlasmaDNAinbloodsamplesof65pregnantwomenwiththegestationalperiodof5〜40weekswereex

5、tractedandprimerextensionpreamplification(PEP)andnestedpolymerasechainreactionwereemployedtoamplifytheSRYgene・Chorionictissue/amnioticcellswereextractedtocomparetheconcordancebetweentheexaminationresultofthemctcrnalbloodandthatofchorionictissue/amnioticcells.Venousbloodofhealthymen

6、andunpregnantwomenwereusedascontrols.Results(1)Thedetectionrateofsinglenucleatedredbloodcellswas90.20%(46/51).(2)TheconcordancerateofSRYgeneamplificationresultsofsinglecellswithrealfetalsexwas82.61%(38/46)、thesensitivityratewasamplificationresultsoffreeDNAwithrealfetalsexwas90.77%(

7、59/65)、thesensitivityratewas89.13%(41/46),andthespecificityratewas94.74%(18/19).Conclusion(1)ThesinglenucleatedredbloodcellsandfreeDNAinmaternalbloodareoffetaloriginandcanbeoneofthevaluablematerialsourcesforprenataldiagnosis・(2)ThedetectionpurityofFNRBCbyusingmicromanipulationtechn

8、iquesisnearly100%anditprov