Abstract
ABSTRACTA forensic validation study of the Early Access Huaxia™ Platinum Polymerase Chain Reaction (PCR) kit was completed to document the performance capabilities and limitations. The genotyping of DNA samples was consistent across a large range of template DNA concentrations, with complete profiles obtained at 0.125 ng; however, no more than 2 mm × 1.2 mm punches of samples would be recommended for direct amplification. The size precision and accuracy test revealed the genotyping ability; while consistent results were obtained when comparing the kit with other commercially available systems. In addition, the whole PCR amplification can finish within approximately 45 min, making the system suitable for fast-detection. However, only partial profiles may be obtained with challenging samples, including DNA stored on Foam-Tipped Applicators (FTA) cards or some case samples. For the forensic application in ethnic groups, a total of 282 and 229 alleles were obtained in Han and Mongolia, respectively. Since the 23 short tandem repeats were independent from each other, the cumulative power of exclusion in duos was 0.999 999 157 188 and the cumulative power of exclusion in trios was 0.999 999 999 859 in the Han group while the cumulative power of exclusion in duos (CPEduo) was 0.999 998 848 26 and cumulative power of exclusion in trios (CPEtrio) was 0.999 999 999 79 in the Mongolia group. And good internal consistency was found between the two investigated groups and the Sichuan Han, Hui, Tibetan and Uygur according to available reference data.
Highlights
Short tandem repeats (STRs) have been the most common genetic markers used in forensic DNA analysis for the past 20 years because the multi-allelic nature of STRs produce many possible genotype combinations among individuals [1,2]
The main experiments were conducted at the Forensic Genetics Laboratory of the Academy of Forensic Science, which is an accredited laboratory by ISO 17025, in accordance with quality control measures
The above results demonstrate that genotypes generated with the Early Access HuaxiaTM Platinum Polymerase Chain Reaction (PCR) kit have been reproducible between laboratories and concordant with the existing STR-amplification systems
Summary
Short tandem repeats (STRs) have been the most common genetic markers used in forensic DNA analysis for the past 20 years because the multi-allelic nature of STRs produce many possible genotype combinations among individuals [1,2]. The Early Access HuaxiaTM Platinum Polymerase Chain Reaction (PCR) kit, which can simultaneously amplify the above mentioned 20 CODIS loci as well as D6S1043 (specially selected for the Chinese population) [4], Penta E Han ethnic group) [5] and Penta D (allele number >10 in the Han ethnic group) [5] were specially designed for the Chinese population by Thermo Fisher Scientific for the application of forensic parentage testing. To evaluate the actual forensic efficiency of the Early Access HuaxiaTM Platinum PCR kit, forensic validation tests, including sensitivity, reliability and repeatability, sizing precision and accuracy, stutter analysis, case sample study and population investigation, were performed. The results obtained and reported here illustrate the performance capabilities and limitations of the multiplex system and how to obtain reliable results required for forensic casework and/or database analysis
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