Y-STR四色荧光复合扩增系统的建立及其应用

Y-STRs Multiplex Amplification and Validation for Forensic Scieuce

目的建立Y染色体STR的四色荧光复合扩增系统,调查7个Y-STR基因座单倍型分布情况。方法设计3套公共引物对分别嵌合在3组Y-STR基因座的原始引物上(1)YS434、Y-GATA-A10、DYS531、DYS557、 DYS448、DYS456、DYS444),再利用加不同荧光颜色标记的3组公共引物对同时复合扩增,用AB I 310遗传分析仪对扩增后产物进行检测,CeneScan、Genotyper软件进行基因分型。结果 3组复合扩增均可成功进行分型,在成都汉族 120名无关男性个体中,7个基因座分别检出4、5、5、8、8、6、7个等位基因,共检出101种单倍型,其中89种为唯一的, 单倍型基因多样性为0．9958。对1例混合斑物证检材,成功检出了与嫌疑人血样Y-STR基因型一致的结果。结论 Y-STR四色荧光标记复合扩增系统分型可靠,对建立Y染色体STR数据库、研究群体遗传学和进行法医学鉴定有重要意义。

Objective In order to increase significantly the discriminatory potential of Y-STR systems available to the forensic community , We have developed and validated a four-color fluorescence multiplex PCR system consisting of 7 male-specific and polymorphic Y-STRs. Methods We designed three sets of constant primers chimerited into three groups of 7 Y-STR＇s specific primers and utilized fluorescence-labeled these three sets of constant primers to amplified simultaneously 7 Y-STRs in a reaction tube. Allele and haplotype frequencies at these Y-STRs were screened by ABI PRISM310 Genetic Analyzer and analyzed by Genotyper software. Results Following optimization of the polymerase chain reaction , DYSCI34, Y-GATA- A10,DYS531 ,DYS557 , DYSCI56 , DS444. and DYS448 in a sample of 120 unrelated males , showed 4, 5, 5, 8, 8, 6,7 alleles respectively. A total of 101 different haplotypes was identified,of which 89 （88. 11% ） were found in single individuals. The overall haplotypes diversity reached 0. 9958. To the one case of mixture stains, our multiplex system drawn conforming conclusion comparing to the result of Y-STR genotypes in suspect＇s blood sample. Conclusion Our results show that the multiplex system of 7 Y-STR will be very powerful for establishing Y-STR database, the paternity testing and mixture stains identifying.