Developmental validation of a 6-dye typing system with 27 loci and application in Han population of China

Yaju Liu,Zheng Li,Meisen Shi,Lihong Guo,Shuhua Ma,Haiying Jin,Rufeng Bai

doi:10.1038/s41598-017-04548-1

Abstract

In this study, a novel 27-locus system (now known as the SureID PanGlobal system), including 24 autosomal STRs (D3S1358, TH01, D21S11, D18S51, Penta E, D12S391, D6S1043, D2S1338, D1S1656, D2S441, D5S818, D13S317, D7S820, D19S433, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D16S539, D22S1045, SE33, D10S1248), two Y-chromosome markers (DYS391 and Y-indel) and the sex determining marker, Amelogenin was developed with six fluorescent dyes labeling. The included STR loci belonged to the core loci in the Combined DNA Index System (CODIS) and the European Standard Set (ESS) as well as some additional loci commonly used in commercial kits and national DNA databases. This paper describes the validation studies conducted with the SureID PanGlobal system using Applied Biosystems 3500 XL Genetic Analyzer for fragment length detection that included the analysis of the following parameters and aspects: PCR conditions, sensitivity, species specificity, inhibition, precision, stutter, DNA mixtures, and stability studies with crime scene samples. The studies demonstrated that the SureID PanGlobal system is reproducible, accurate, sensitive and robust for forensic application and databasing. Additionally, the whole cycling time of the system can finish within 65 minutes, which was developed specifically for rapid and reliable generation of DNA profiles obtained from blood, buccal swabs and forensic stains.

Highlights

Short tandem repeats (STRs) typing methods are widely used for paternity testing, human identification, and DNA database searches in criminal investigations1, 2
Annealing temperatures between 56 °C and 66.5 °C were tested in triplicates with 1.0 ng Control DNA 9948
The whole PCR reaction can finish within 65 min, which was developed for rapid and reliable generation of DNA profiles obtained from blood, buccal swabs and forensic stains

Summary

Introduction

Short tandem repeats (STRs) typing methods are widely used for paternity testing, human identification, and DNA database searches in criminal investigations . As DNA databases continue to grow and international cooperation increases, there is still a need to develop sufficient multiplex systems to facilitate data sharing and to minimize adventitious matches In this sense, we developed a 6-dye typing system ( known as the SureID PanGlobal Human DNA Identification Kit) consisting of 24 autosomal STRs which combined both CODIS and ESS core STR loci (CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, D22S1045, D2S441, D1S1656, and D12S391), 7 other routinely tested loci (Penta D, Penta E, D2S1338, D19S433, D6S1043, D10S1248, and SE33) included to increase the discriminatory power of the multiplex, plus amelogenin (AMEL) for sex discrimination and DYS391, Yindel, the last loci incorporated to prevent improper gender determination when null Y alleles or deletions of the Y-chromosome may involve the locus amelogenin. The conclusions reported here support the fact that the 6-dye SureID PanGlobal system is suitable for database applications and human identification in casework

Methods

Results

Conclusion