Developmental Validation of the Huaxia Platinum System and application in 3 main ethnic groups of China.

Zheng Wang,Chengtao Li,Luyao Li,Wei Wu,Yiping Hou,Zhenjun Jia,Di Zhou

doi:10.1038/srep31075

Abstract

STRs, scattered throughout the genome with higher mutation rate, are attractive to genetic application like forensic, anthropological and population genetics studies. STR profiling has now been applied in various aspects of human identification in forensic investigations. This work described the developmental validation of a novel and universal assay, the Huaxia Platinum System, which amplifies all markers in the expanded CODIS core loci and the Chinese National Database in one single PCR system. Developmental validation demonstrated that this novel assay is accurate, sensitive, reproducible and robust. No discordant calls were observed between the Huaxia Platinum System and other STR systems. Full genotypes could be achieved even with 250 pg of human DNA. Additionally, 402 unrelated individuals from 3 main ethnic groups of China (Han, Uygur and Tibetan) were genotyped to investigate the effectiveness of this novel assay. The CMP were 2.3094 × 10−27, 4.3791 × 10−28 and 6.9118 × 10−27, respectively, and the CPE were 0.99999999939059, 0.99999999989653 and 0.99999999976386, respectively. Aforementioned results suggested that the Huaxia Platinum System is polymorphic and informative, which provides efficient tool for national DNA database and facilitate international data sharing.

Highlights

Short tandem repeats (STRs) are DNA regions with a variable number of tandemly repeated short (2–6 bp) sequence motifs[1]
The results showed that full profiles were obtained except at 0.5×master mix volume
These results demonstrated that this novel assay could tolerate moderate deviations to the recommended concentration of primer set and master mix

Summary

Introduction

Short tandem repeats (STRs) are DNA regions with a variable number of tandemly repeated short (2–6 bp) sequence motifs[1]. Multiplex that allows co-amplification and fluorescent detection of the 23 loci (CSF1PO, FGA, TH01, TPOX, vWA, D1S1656, D2S1338, D2S441, D3S1358, D5S818, D7S820, D8S1179, D10S1248, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11, D22S1045, D6S1043, Penta D and Penta E) including all the recommended core loci in the expanded CODIS core loci and the Chinese National Database as well as Amelogenin and Y-InDel (rs2032678) for gender determination (More detailed information can be found in Supplementary Table S2) It has a new improved PCR buffer formulation and optimized PCR cycling conditions (less than 50 minutes) to enable direct amplification of blood samples. The validation results demonstrated that the Huaxia Platinum System is robust, specific, sensitive, stable and reliable for human identification applications

Methods

Results

Conclusion