目的基于二代测序技术(next generation sequencing, NGS)用于Y染色体多拷贝短串联重复序列(multi-copy short tandem repeat on Y chromosome, Multi-copy Y-STRs)DYF404S1分型方法建立及在河南汉族群体中遗传多态性分析的初步研究。...目的基于二代测序技术(next generation sequencing, NGS)用于Y染色体多拷贝短串联重复序列(multi-copy short tandem repeat on Y chromosome, Multi-copy Y-STRs)DYF404S1分型方法建立及在河南汉族群体中遗传多态性分析的初步研究。方法在DYF404S1基因座特异引物两端添加适宜的接头,得到带有不同接头的引物组合,共25组。采用BIOO RAPID DNA试剂盒构建二代测序文库,使用Illumina MiseqTM二代测序仪对225例河南汉族群体的男性样本进行测序,STRait Razor V2.0软件进行测序数据分析,同时与一代测序ABI3100遗传分析仪分型结果比对分析。结果基于Illumina MiseqTM二代测序平台,采用引物两端加接头的方法对225例河南汉族无关男性个体样本进行DYF404S1基因座分型,其中220例成功分型,成功率达97.8%;同时使用3130遗传分析仪进行DYF404S1基因座分型,分型结果与二代测序分型结果一致。3例样本出现等位基因异常分型,均表现为3等位基因模式。DYF404S1基因座在河南汉族群体中的单体型多样性(haplotype diversity, HD)和个体识别力(discrimination power, DP)分别为0.9356和0.9311。结论初步建立了NGS技术用于多拷贝DYF404S1分型方法,为法医学实践和群体遗传学提供了技术和理论基础。展开更多
Two rare cases of long alleles at Y‑chromosome short tandem repeat(Y‑STR)loci(DYF387S1 and DYS447)were identified when two father-son pairs were analyzed by multiplex amplification.“Null alleles”were observed at DYF...Two rare cases of long alleles at Y‑chromosome short tandem repeat(Y‑STR)loci(DYF387S1 and DYS447)were identified when two father-son pairs were analyzed by multiplex amplification.“Null alleles”were observed at DYF387S1 and DYS447,and duplicated alleles were displayed at DYS533 and DYS19.We secondly amplified DYF387S1,DYS533,DYS447,and DYS19 loci by singleplex polymerase chain reaction(PCR)and sequence analysis of the long alleles at DYF387S1 and DYS447 loci.The results showed that alleles from DYF387S1(allele 55)and DYS447(allele 41)were longer than their common sizes in the allelic ladder ranges(33-42 for DYF387S1 and 18-30 for DYS447)and located in the neighboring loci(DYS533 and DYS19,respectively).Therefore,to identify these cases involving this unusual phenomenon,not only re‑amplification using the same kit but also additional amplification(using alternative multiplex kits with different adjoining markers or additional singleplex PCR amplification)should be performed to avoid misinterpreting Y‑STR profiles.展开更多
文摘Two rare cases of long alleles at Y‑chromosome short tandem repeat(Y‑STR)loci(DYF387S1 and DYS447)were identified when two father-son pairs were analyzed by multiplex amplification.“Null alleles”were observed at DYF387S1 and DYS447,and duplicated alleles were displayed at DYS533 and DYS19.We secondly amplified DYF387S1,DYS533,DYS447,and DYS19 loci by singleplex polymerase chain reaction(PCR)and sequence analysis of the long alleles at DYF387S1 and DYS447 loci.The results showed that alleles from DYF387S1(allele 55)and DYS447(allele 41)were longer than their common sizes in the allelic ladder ranges(33-42 for DYF387S1 and 18-30 for DYS447)and located in the neighboring loci(DYS533 and DYS19,respectively).Therefore,to identify these cases involving this unusual phenomenon,not only re‑amplification using the same kit but also additional amplification(using alternative multiplex kits with different adjoining markers or additional singleplex PCR amplification)should be performed to avoid misinterpreting Y‑STR profiles.
