| ORIGINAL ARTICLE |
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| Year : 2015 | Volume
: 1
| Issue : 1 | Page : 8-15 |
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Validation of the DNATyper™15 PCR Genotyping System for Forensic Application
Jian Ye1, Chengtao Jiang2, Xingchun Zhao1, Le Wang1, Caixia Li1, Anquan Ji3, Li Yuan4, Jing Sun1, Shuaifeng Chen5
1 Institute of Forensic Science, Ministry of Public Security; Beijing Engineering Research Center of Crime Scene Evidence Examination; Key Laboratory of Forensic Genetics, Ministry of Public Security, Beijing, China
2 Fada Institute of Forensic Medicine and Science, Beijing, China
3 Institute of Forensic Science, Ministry of Public Security, Beijing, China
4 Fada Institute of Forensic Medicine and Science; Key Laboratory of Evidence Science, China University of Political Science and Law, Ministry of Education, Beijing, China
5 Department of Investigation, People's Public Security University of China, Beijing, China
Correspondence Address:
Jian Ye
Beijing No. 3817 Letterbox, Beijing - 100038
China
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/2349-5014.157903
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We describe the optimization and validation of the DNATyper™15 multiplex polymerase chain reaction (PCR) genotyping system for autosomal short tandem repeat (STR) amplification at 14 autosomal loci (D6S1043, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, D13S317, D8S1179, D16S539, Penta E, D5S818, vWA, D18S51, and FGA) and amelogenin, a sex-determining locus. Several DNATyper™15 assay variables were optimized, including hot start Taq polymerase concentration, Taq polymerase activation time, magnesium concentration, primer concentration, annealing temperature, reaction volume, and cycle number. The performance of the assay was validated with respect to species specificity, sensitivity to template concentration, stability, accuracy, influence of the DNA extraction methods, and the ability to genotype the mixture samples. The performance of the DNATyper™15 system on casework samples was compared with that of two widely used STR amplification kits, Identifiler™ (Applied Biosystems, Carlsbad, CA, USA) and PowerPlex 16 ® (Promega, Madison, WI, USA). The conditions for PCR-based DNATyper™15 genotyping were optimized. Contamination from forensically relevant nonhuman DNA was not found to impact genotyping results, and full profiles were generated for all the reactions containing ≥ 0.125 ng of DNA template. No significant difference in performance was observed even after the DNATyper™15 assay components were subjected to 20 freeze-thaw cycles. The performances of DNATyper™15, Identifiler™, and PowerPlex 16 ® were comparable in terms of sensitivity and the ability to genotype the mixed samples and case-type samples, with the assays giving the same genotyping results for all the shared loci. The DNA extraction methods did not affect the performance of any of the systems. Our results demonstrate that the DNATyper™15 system is suitable for genotyping in both forensic DNA database work and case-type samples. |
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