Deng Zhi-Hui, Li Qian, Wu Shuang, Li Da-Cheng, Yang Bao-Cheng
Shenzhen Institute of Transfusion Medicine, Shenzhen, 518035, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Jun;16(3):699-703.
The purpose of this study was to explore the ability of discrimination of the AmpFlSTR Yfiler PCR amplification kit containing 17 Y-STR loci and the allelic mutation in the practice of paternity testing in Chinese population. 36 non-paternity father/son pairs and 84 confirmed father/son pairs, which had been previously genotyped by using Reliagene Y-PLEX 6 commercial kit and the "9 Y-STR multiplex with reduced-size amplicons" developed by our laboratory, were subjected to Y-STR genotyping at 17 loci using the AmpFlSTR Yfiler PCR amplification kit. 17 Y-STR loci were amplified in single multiplex and the PCR products were detected by using ABI Prism 3100 DNA Sequencer. The number of Y-STR exclusion for each non-paternity father/son pair and the mutation events for each confirmed father/son pair were calculated and the observed results were compared with our previous reported data determined by Reliagene Y-PLEX 6 kit and the "9 Y-STR multiplex with reduced-size amplicons". The results showed that out of 36 non-paternity father/son pairs subjected to Y-STR genotyping by using the AmpFlSTR Yfiler kit, one case with no Y-STR exclusion of paternity and 35 cases with more than 3 Y-STR exclusions for each father/son pair were observed. The percentage of cases with more than 3 Y-STR exclusions in all the tested non-paternity cases for Yfiler kit was 97.22% (35/36), which was more than that of Reliagene Y-PLEX 6 kit (92.11%, 35/38) and our "9 Y-STR multiplex with reduced-size amplicons" (91.67%, 33/36). Except for single father/son pair with no Y-STR exclusion, an average of 11.3 Y-STR exclusions was observed in other 35 non-paternity father/son pairs. In the 84 confirmed father/son pairs, 5 mutation events with a single unit repeat change at DYS437, DYS439, DYS635, DYS389II and DYS19, respectively, were identified using the Yfiler kit. The average mutation rate was estimated at 3.50 x 10(-3) per locus per generation. The cases with Y-STR mutation events in all tested confirmed father/son pairs for Yfiler system were 5.95% (5/84), which was significantly higher than that of Y-PLEX 6 (2.15%, 2/93) and "9 Y-STR multiplex with reduced-size amplicons" (no mutation events in the same 84 confirmed father/son pairs). It is concluded that the Yfiler kit which allowing simultaneous analysis of 17 Y-STR loci offers a high ability of discrimination for paternity testing, however, the Y-STR allelic mutation of the Yfiler system can not be neglected.
本研究旨在探讨包含17个Y-STR基因座的AmpFlSTR Yfiler PCR扩增试剂盒在中国人群亲子鉴定实践中的鉴别能力及等位基因突变情况。36对非亲生父子对和84对已确认的亲生父子对，此前曾使用Reliagene Y-PLEX 6商业试剂盒及本实验室研发的“9个Y-STR复合扩增体系（扩增子片段减小）”进行基因分型，现使用AmpFlSTR Yfiler PCR扩增试剂盒对这17个基因座进行Y-STR基因分型。17个Y-STR基因座在单个复合扩增体系中进行扩增，PCR产物使用ABI Prism 3100 DNA测序仪进行检测。计算每对非亲生父子对的Y-STR排除数以及每对已确认亲生父子对的突变事件数，并将观察结果与我们先前使用Reliagene Y-PLEX 6试剂盒及“9个Y-STR复合扩增体系（扩增子片段减小）”所报告的数据进行比较。结果显示，在使用AmpFlSTR Yfiler试剂盒进行Y-STR基因分型的36对非亲生父子对中，观察到1例无Y-STR排除亲子关系的情况，以及35例每对父子对有超过3个Y-STR排除的情况。在所有使用Yfiler试剂盒检测的非亲生案例中，有超过3个Y-STR排除的案例百分比为97.22%（35/36），高于Reliagene Y-PLEX 6试剂盒（92.11%，35/38）和我们的“9个Y-STR复合扩增体系（扩增子片段减小）”（91.67%，33/36）。除1对无Y-STR排除的父子对外，在其他35对非亲生父子对中平均观察到11.3个Y-STR排除。在84对已确认的亲生父子对中，使用Yfiler试剂盒鉴定出5例突变事件，分别在DYS437、DYS439、DYS635、DYS389II和DYS19处发生单个单位重复改变。估计平均突变率为每基因座每代3.50×10⁻³。在所有使用Yfiler系统检测的已确认亲生父子对中，发生Y-STR突变事件的案例为5.95%（5/84），显著高于Y-PLEX 6（2.15%，2/93）和“9个Y-STR复合扩增体系（扩增子片段减小）”（在相同的84对已确认亲生父子对中无突变事件）。结论是，可同时分析17个Y-STR基因座的Yfiler试剂盒在亲子鉴定中具有较高的鉴别能力，然而，Yfiler系统的Y-STR等位基因突变不容忽视。
