用于犬类生物材料法医DNA分析的短串联重复序列试剂盒的发育验证
Developmental validation of short tandem repeat reagent kit for forensic DNA profiling of canine biological material.
作者信息
Dayton Melody, Koskinen Mikko T, Tom Bradley K, Mattila Anna-Maria, Johnston Eric, Halverson Joy, Fantin Dennis, DeNise Sue, Budowle Bruce, Smith David Glenn, Kanthaswamy Sree
机构信息
Department of Anthropology, University of California, One Shields Avenue, Davis, CA 95616, USA.
出版信息
Croat Med J. 2009 Jun;50(3):268-85. doi: 10.3325/cmj.2009.50.268.
AIM
To develop a reagent kit that enables multiplex polymerase chain reaction (PCR) amplification of 18 short tandem repeats (STR) and the canine sex-determining Zinc Finger marker.
METHODS
Validation studies to determine the robustness and reliability in forensic DNA typing of this multiplex assay included sensitivity testing, reproducibility studies, intra- and inter-locus color balance studies, annealing temperature and cycle number studies, peak height ratio determination, characterization of artifacts such as stutter percentages and dye blobs, mixture analyses, species-specificity, case type samples analyses and population studies.
RESULTS
The kit robustly amplified domesticated dog samples and consistently generated full 19-locus profiles from as little as 125 pg of dog DNA. In addition, wolf DNA samples could be analyzed with the kit.
CONCLUSION
The kit, which produces robust, reliable, and reproducible results, will be made available for the forensic research community after modifications based on this study's evaluation to comply with the quality standards expected for forensic casework.
目的
开发一种试剂盒，能够对18个短串联重复序列（STR）和犬类性别决定锌指标记进行多重聚合酶链反应（PCR）扩增。
方法
为确定该多重检测方法在法医DNA分型中的稳健性和可靠性而进行的验证研究包括灵敏度测试、重复性研究、位点内和位点间颜色平衡研究、退火温度和循环次数研究、峰高比测定、诸如拖尾百分比和染料斑点等假象的特征分析、混合分析、物种特异性、案件类型样本分析和群体研究。
结果
该试剂盒能有力地扩增家养犬样本，并始终能从低至125 pg的犬类DNA中生成完整的19个位点图谱。此外，该试剂盒还可用于分析狼的DNA样本。
结论
该试剂盒产生的结果稳健、可靠且可重复，在根据本研究的评估进行修改以符合法医案件工作预期的质量标准后，将提供给法医研究界。
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