12个mini-X-STR和Amel基因座荧光标记复合扩增体系的初步研究

Development of a multiplex PCR system for analysis of 12 mini-X-chromosomal STR loci and Amelogenin gene

目的研究建立12个mini-X-STR和Amel基因座复合扩增体系。方法选择DXS101、DXS10159、DXS10162、DXS10164、DXS6789、DXS7133、DXS7423、DXS7424、DXS8378、DXS981、GATA165B12和GA-TA31E08共12个X-STR与Amelogenin基因座,自行设计引物,分别采用FAM、HEX、Tamra、ROX四色荧光标记5′端,并进行必要的修饰,按照合适的引物浓度进行混合。对97个血痕样本进行复合扩增,并用ABI3130XL进行电泳和分型。结果所建立的荧光标记复合扩增体系能够得到清晰、明确的分型图谱,灵敏度达50pg,稳定性、重复性与平衡性较佳。结论本研究建立的12个mini-X-STR和Amel基因座荧光标记复合扩增体系统检验方法简单,灵敏度高,适合实际办案的需要。

Objective To develop a 12 mini-X-chromosomal STR multiplex PCR system. Methods DNA were extracted from bloodstains. Primers of 12 X-chromosomal STRs （DXS7133, DXS8378, DXS981, DXS7424, DXS6789, DXS10159, GA- TAJ65B12, DXS101, DXS7423, DATA31E08, DXS10164, DXS10162） and Amelogenin gene were labeled with four different fluorescent dyes： FAM, HEX, TAMRA and Rox. PCR were performed in a single reaction. PCR products of 97 bloodstains samples were separated on Genetic analyzer 3130XL and STRs were typed by using GeneMapper ID 3.2. Results All of 97 samples have a clear and unambiguous typing, although mini-stutters were observed in a few loci. The same typing could be gained when samples were typed repeatedly. The sensitive of this multiplex PCR was low to 50pg. Conclusion Established multiplex PCR system for 12 mini-X-chromosomal STR loci and amelogenin gene can be used in forensic practice.