常染色体短串联重复序列基因座复合扩增等位基因丢失现象的研究  

Analysis of allelic drop-out at autosomal short tandem repeat loci in composite amplification

作　　者：高素青[1] 金士正[1] 何柳媚[1] 李桢[1] 邓志辉[1] 洪文旭[1] Gao Suqing;Jin Shizheng;He Liumei;Li Zhen;Deng Zhihui;Hong Wenxu(Shenzhen Blood Center Institute,Shenzhen 518035,Guangdong Province,China)

机构地区：[1]广东省深圳市血液中心研究所,518035

出　　处：《国际输血及血液学杂志》2018年第5期397-405,共9页International Journal of Blood Transfusion and Hematology

基　　金：深圳市卫生计生系统科研项目（201606046）

摘　　要：目的探讨常染色体短串联重复序列（STR）基因座复合扩增等位基因丢失现象的特征和原因，以及亲子鉴定中STR基因座等位基因疑似突变的确认策略。方法选择2017年1月至6月，于深圳市血液中心接受三联体亲子鉴定的3个家庭的9例家庭成员为研究对象。研究对象纳入标准：STR基因座等位基因初检结果不符合孟德尔遗传规律，疑为STR基因座等位基因丢失者。采用美国ABI公司生产的GlobalFiler Express试剂盒对受试者血斑片样本的21个常染色体STR基因座等位基因进行分型。采用美国Promega公司生产的PowerPlex 21复合扩增系统对受试者血斑片样本的STR等位基因分型结果进行验证。采用PCR-直接测序（SBT）法，检测受试者DNA样本人类白细胞抗原（HLA） -A/-B/-C/-DR/-DQB1基因座等位基因，并且对受试者HLA基因座单倍型进行分型，作为个体识别的补充验证。本研究所遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》，并且与受试者或其监护人签订知情同意书。结果①本研究3个家庭的三联体亲子鉴定中，6例受试者丢失的等位基因分别发生在STR的3个基因座CSF1PO、D1S165和TH01，并且突变步数均不相同。上述丢失的等位基因，既可能为父源性，亦可能为母源性。②家庭1中，对母亲和孩子的CSF1PO基因座等位基因分型的初检结果显示，等位基因均为纯合子；验证结果显示，等位基因均为杂合子；二者的CSF1PO基因座等位基因初检结果均丢失等位基因10。③家庭2中，对父亲和孩子的D1S1656基因座等位基因分型的初检结果显示，等位基因均为纯合子；验证结果显示，等位基因均为杂合子；二者的D1S1656基因座等位基因初检结果均丢失等位基因16。④家庭3中，对母亲和孩子的TH01基因座等位基因分型的初检结果显示，等位基因均为纯合子；验证结果显示，等位基因均为杂ObjectiveTo investigate the characteristics and causes of allelic drop-out at the autosomal short tandem repeat （STR） loci, and to explore the confirmation strategy of suspected mutations at the STR loci alleles in paternity.MethodsFrom January to June 2017, nine family members from 3 families of three-banded patterns at Shenzhen Blood Center were selected as subjects. Subject inclusion criteria： subjects′ initial test results of the STR loci alleles did not conform to the Mendels laws of genetics, which were suspected allelic drop-out at STR loci. GlobalFiler？ Express kits from ABI, USA were initially used to detect genotype of 21 autosomal STR loci alleles from all blood spot samples. All blood spot samples suspected for having allelic drop-out were verified with the PowerPlex？ 21 kits from Promega, USA. Supplemental experiment was used human leukocyte antigen （HLA）-A/-B/-C/-DR/-DQB1 loci genotype test results with PCR-sequence-based typing （SBT） method, and then the haplotype of the subjects was deduced by the family analysis to complete the individual identification verification. This study met the requirements of World Medical Association Declaration of Helsinki revised in 2013. Informed consent was obtained from every subject. Results① In this study, the allelic drop-out in 6 subjects occurred in 3 STR loci of CSF1PO, D1S165 and TH01, respectively, and the number of mutation steps were different. These allelic drop-out could be either paternal or maternal. ② In family 1, the initial test result of CSF1PO locus was homozygous in both mother and child. The verification test result of CSF1PO locus was heterozygous, and allelic drop 10-locus were confirmed. ③ In family 2, the initial test result of D1S165 locus was homozygous in both child and father. The verification test result of D1S165 locus was heterozygous, and allelic drop-out 16-locus was confirmed. ④ In family 3, the initial test result of TH01 locus was homozygous in both the mother and child. The verification test result

关 键 词：杂合子丢失 等位基因 串联重复序列 HLA抗原 亲子鉴定 

分 类 号：D919[医药卫生—法医学]