To develop and validate a novel 6-dye STR(short tandem repeat)25-plex DNA typing system for forensic DNA profiling and databases.In this study,a novel STR 25-plex DNA typing system that includes 24 autosomal STRs(D1S1656,D2S1338,D2S441,D3S1358,D5S818,D6S1043,D7S820,D8S1179,D10S1248,D12S391,D13S317,D16S539,D18S51,D19S433,D21S11,D22S1045,CSF1PO,FGA,Penta D,Penta E,TH01,TPOX,vWA,D11S4463)and Amelogenin was developed.Validation studies demonstrated the sensitivity,accuracy,and reproducibility of our novel STR 25-plex DNA typing system.The sensitivity of the STR 25-plex DNA typing system was demonstrated by the ability to obtain complete profiles from as little as 0.125ng of human DNA.Specificity testing was demonstrated by the lack of cross-reactivity to a variety of commonly encountered animal species and microbial pool.For stability testing,full profiles were obtained with humic acid concentration≤60ng/μL and hematin≤600μM.For forensic evaluation,the selected 24 autosomal STRs followed the Hardy-Weinberg equilibrium.Since 24 autosomal STRs were independent from one another,PM(Probability matching)was 3.5434×10-28,TDP(Total Probability of Discrimination Power)was 0.999999999999999999999999969863,and CEP(Cumulative probability of exclusion)was 0.99999999375.The new STR 25-plex typing system is sensitive,reproducible,and stable,therefore it is highly applicable for use in national DNA database and can help to facilitate international data sharing.
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