摘要
目的对一例疑似串联重复基因座出现三带型的新生儿进行分析,明确其准确的基因分型并对其特点进行分析。方法采集新生儿足底血,初次检验采用华夏TM白金PCR扩增试剂盒进行STR基因座检测分型,随后采用GoldeneyeTMDNA身份鉴定系统25A试剂盒进行复检。同时采用单基因座PCR扩增结合聚丙烯酰胺凝胶电泳分型技术分离单一条带进行测序验证。结果初检D12S391基因座出现三带型,基因分型为13/17/19。其余基因座基因分型未见异常,其中D10S1248基因座分型为13/16,与D12S391相邻的基因座D1S1656基因分型为纯合子16。复检后D12S391基因座分型为17/19,D1S1656基因座分型仍为纯合子16,与D1S1656相邻的D10S1248基因座分型为13/16,同时出现一个未被标识的OL峰。推测初检时在D12S391基因座出现的等位基因13和复检时在D10S1248出现的OL峰均为D1S1656基因座的稀有等位基因22。D1S1656基因座出现假纯合子,实际基因分型应为16/22且经测序证实。结论确定了该例新生儿STR基因座出现的三等位基因为假三带型。对于异常三等位基因,需更换其他试剂盒进行复核验证,必要时可进行单基因座扩增测序,从而保证基因分型的准确性、科学性和可靠性。
Objective Analysis of the suspected three-banded patterns at short tandem repeats in a newborn to obtain the accurate genotyping and analyse its characteristics.Methods Blood sample were collected from the feet of the newborn.In the first,the genotypes of STR loci were detected by Huaxia TM platinum PCR kit,then the GoldeneyeTM DNA ID System 25 A was used to perform the genotyping once again.Meanwhile single allele band separated by singleplex PCR combining the polyacrylamide gel electrophoresis was sequenced.Results In the first trial,D12 S391 showed the three-banded patterns and the genotype was 13/17/19,the rest loci were normal with the genotype 13/16 at D10 S1248 and the homozygote 16 at D1 S1656 that is adjacent to the D12 S391.In the second time,the genotype was 17/19 at D12 S391 and D1 S1656 showed the homozygote 16.D10 S1248 that is close to the D1 S1656 showed the genotype of 13/16,but an unidentified allele peak that was named OL was detected.It is speculated that both the allele 13 at D12 S391 in the first trial and the unidentified allele peak OL in the second time is the rare allele 22 belonged to D1 S1656.The genotype of D1 S1656 should be heterozygous genotype 16/22 not homozygous genotype,which was validated by Sanger sequencing.Conclusion It was confirmed that the tri-alleles at STR locus in this case was false three-banded patterns.For the abnormal tri-alleles in STR loci,it is necessary to replace the kit for verification,and if necessary,single locus amplification and sequencing can be carried out to ensure the accuracy,scientificity and reliability of the genotype.
作者
刘奇
张国安
李书越
虢洪松
韩亚文
侯秀迪
孙倩倩
王业全
LIU Qi;ZHANG Guoan;LI Shuyue;GUO Hongsong;HAN Yawen;HOU Xiudi;SUN Qianqian;WANG Yequan(Institute of Forensic Medicine and Laboratory Medicine,Jining Medical University,Jining,Shandong 272067,China;Forensic Science Center of Jining Medical University,Jining,Shandong 272013,China)
出处
《中国优生与遗传杂志》
2021年第2期243-245,共3页
Chinese Journal of Birth Health & Heredity
基金
济宁医学院教师科研扶持基金(JYFC2018FY004,JYFC2018FY001)。
