DNATyper^(TM) 30六色荧光STR复合扩增冻干试剂的验证  

Comparation of Lyophilized and Normal DNATyper^(TM) 306-dye PCR Amplifi cation Kit

作　　者：张哲 于浩 李晨佳 余政梁 吴俞衡 张广峰 李万水 白雪 ZHANG Zhe;YU Hao;LI Chenjia;YU Zhengliang;WU Yuheng;ZHANG Guangfeng;LI Wanshui;BAI Xue(Institute of Forensic Science,Ministry of Pubic Security,Beijing 100038,China;Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]公安部鉴定中心,北京100038 [2]河北医科大学,石家庄050017

出　　处：《刑事技术》2023年第6期616-622,共7页Forensic Science and Technology

基　　金：公安部科技强警基础工作专项(2021JC13);国家重点研发计划(2021YFC3300104)。

摘　　要：本研究旨在测试DNATyper^(TM) 30冻干试剂的技术性能指标,评估其在法医学案件检验中的应用能力。制定测试方案,设置不同的存储条件和时间范围,通过检测标准DNA 9947A对冻干试剂的检出率、灵敏度、稳定性等方面进行测试,并使用案件样本对冻干试剂进行检测,同时将液体试剂和冻干试剂性能进行平行比对。结果显示,DNATyper^(TM) 30冻干试剂分型结果准确,可重复性好,具有较高的灵敏度(0.125 ng),对微量陈旧检材有很好的适应性,可常温状态下保存1个月以上。本研究表明DNATyper^(TM) 30冻干试剂可显著增加上样模板量、可延长试剂在常温下的保存时间、操作简单、灵敏度高,可满足微量DNA检测、试剂长途运输和常温携带的需要。Freeze drying technology can remove about 99%moisture from the substance,resulting in the freeze-dried product with an increased internal surface area,which is loose,porous,and easy to recover.These characteristics are very suitable for application in the PCR amplification kit of forensic STR examination.Lyophilized reagent could break the challenge of the upper limit of template addition in conventional PCR systems.This study aims to test the technical performance of DNATyper^(TM) 30 lyophilized reagent and evaluate its application ability in practical forensic cases.In order to test the detection rate,sensitivity,stability and other aspects of lyophilized reagents,an exploration was carried out by setting different storage conditions,time ranges and concentration gradient.In order to meet the needs of room temperature transportation,the reagents were placed at room temperature(23°C to 25°C)to simulate the transportation environment and compare the STR typing results at 1,7,15,30 days.In sensitivity test,standard DNA 9947A was used as template and its gradient was set from 0.0313 ng to 2ng.Furthermore,three types of case samples were selected,including trace DNA sample,to test the practical application of lyophilized reagents and liquid reagents were used in parallel for comparison.The results showed that DNATyper^(TM) 30 lyophilized reagent had accurate typing results and the peak value was between 3000 and 20000 RFUs with good repeatability.The lyophilized reagent was validated to get a well-balanced STR typing result,and the sensitivity was 0.125 ng,while it showed good stability when stored at room temperature for over 1 month.Essentially,lyophilized reagents performed outstandingly in the detection of trace DNA samples,with a higher detection rate and higher peak height of loci compared to conventional reagents.Besides,compared to longer segment loci,lyophilized reagents had a higher detection rate for shorter segment loci.Overall,DNATyper^(TM) 30 lyophilized reagent,which is easy to operate and has a hig

关 键 词：法医遗传学 冻干试剂 微量DNA 短串联重复序列 DNATyper^(TM)30试剂盒 

分 类 号：DF795.2[医药卫生—法医学]