为解决司法机关对猫、鸭、牛、羊四种动物组织成分种属的鉴别问题,构建一个同时鉴定猫、鸭、牛、羊4种常见物种的STR基因座复合扩增体系.根据NCBI收录及基因组筛查,猫、鸭、牛和羊经检测各筛查出4个具有种属特异性强和引物扩增效果好的STR基因座,应用Primer premier 5.0软件设计引物,在5'端添加荧光引物FAM、HEX、ROX,TAMRA,对16个基因座进行复合扩增检验.通过实验构建的包含16个STR基因座的复合扩增体系,分型灵敏度高、特异性强且结果准确.FAM等4种荧光引物标记用于猫、鸭、牛、羊的种属鉴定,可为司法机关食药监部门实践及刑事案件中非人组织的种属鉴定提供科学的鉴定依据.
The Construction and Applications of a Composite Amplification System for 16 STR Loci for Species Identification of Cat,Duck,Cow,and Sheep
To address the issue of identifying the species composition of cats,ducks,cows,and sheep by judicial authorities,a compound amplification system for STR loci was developed to simultaneously identify these four common species. Based on data from NCBI and genomic screening,four STR loci with strong species specificity and good primer amplification effects were identified for cats,ducks,cows,and sheep. Using Primer Premier 5.0 software,primers were designed with fluorescent labels FAM,HEX,ROX,and TAMRA added to the 5' end,allowing for multiplex amplification testing of 16 loci. The multiplex amplification system constructed through experimentation,which includes 16 STR loci,demonstrates high typing sensitivity,strong specificity,and accurate results. The four fluorescently labeled primers,including FAM,are used for species identification of cats,ducks,cows,and sheep,providing a scientific basis for species identification of non-human tissues in the practices of judicial food and drug regulatory authorities,as well as in criminal cases.
forensic geneticscomposite amplification systemshort tandem repeats sequencesspecies identificationfood safety
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