Forensic genetic analysis of population of Madhya Pradesh with PowerPlex Fusion 6C™ Multiplex System
Performance of PowerPlex Fusion 6C kit (PP F6C) was assessed in 374 unrelated individuals belonging to Madhya Pradesh, an Indian state. The study evaluated the forensic parameters for the loci included in PP F6C Multiplex System. The combined discrimination power (CPD) and combined exclusion power (CPE) were 1 and 0.999999995, respectively, for all 23 autosomal STR loci. SE33 showed the greatest power of discrimination (0.990) in the studied population, whereas TPOX showed the lowest (0.843). The availability of three Y-STR loci in the Multiplex System is suitable for assessing male contribution and amelogenin deletion in a single Multiplex PCR simultaneously. The study also presents the first global report on polymorphism in the Indian population on SE 33 autosomal STR loci and PP Fusion 6C Multiplex System. The results revealed that the studied STR Multiplex System is highly polymorphic and suitable for forensic purposes.
KeywordsForensic DNA fingerprinting STR Madhya Pradesh India PowerPlex fusion 6C
Present capillary electrophoresis and PCR-based STR analysis using expanded Combined DNA Index System (CODIS) 20 markers are state of art in forensic DNA typing . For performing the identification conclusively and precisely with a significant probability value, an increase in the number of loci is recommended, as the discrimination power of any Multiplex System is based on a combination of all the independent loci tested. Madhya Pradesh is the second largest state in India by area. The population of Madhya Pradesh is 72,597,565 comprising of 37,612,920 males and 34,984,645 females, contributing 6% to India’s total population as per the 2011 census. A few studies have been performed to characterize the population of Madhya Pradesh on autosomal STRs (described in Table S2) but these studies are based on 15 STR markers either using Identifiler/Identifiler Plus (Applied Biosystem, USA) or PowerPlex 16/16HS (Promega Corporation, Madison, USA).
Based on six dyes, PP F6C System (Promega Corporation, Madison, USA) is a 27-locus multiplex that ensures simultaneous detection of 20 autosomal loci in the expanded CODIS (CSF1PO, FGA, TH01, TPOX, vWA, D1S1656, D2S1338, D2S441, D3S1358, D5S818, D7S820, D8S1179, D10S1248, D12S391, D13S317, D16S539, D18S51, D19S433, D21S11, and D22S1045), three more loci for better discrimination (Penta D, Penta E and SE33), Amelogenin for sex determination and DYS391 and two rapidly mutating Y-STR loci, DYS570 and DYS576 for further confirmation of gender. A multi-laboratory evaluation of the PP F6C System has already been published  following SWGDAM guidelines . This study is the first global attempt to work out the efficacy of PP F6C System and the first ever population data statement about SE 33 marker in Indian population for forensic purposes. Samples (n = 374) of unrelated individuals were taken from routine casework analysis performed between January 2018 and October 2018 by the authors who work at the DNA Fingerprinting Unit, State Forensic Science Laboratory, Sagar, Madhya Pradesh, India. The samples included in this study have not been typed by any other Multiplex System and this is the first study from India detailing the forensic efficacy of a 27-marker multi-utility STR Multiplex System. Following the Code of Ethics of the World Medical Association (i.e., Declaration of Helsinki) for experiments involving humans, peripheral blood samples of these unrelated individuals were collected. All individuals provided prior written informed consent. No minor was involved in the study.
After DNA extraction and PCR, analysis was performed on 3500XL genetic analyzer, and statistical evaluation was done as described before . The peak height threshold was 50 RFU for heterozygous and 200 RFU for homozygous alleles. Internal laboratory control standards and kit controls were used at all steps of analysis. The authors have passed the proficiency test of the GITAD, Spain (http://gitad.ugr.es/principal.htm). This article follows the population data publication guidelines formulated by the journal.
The allele frequencies and results of the forensic efficiency parameters for the 23 autosomal STR loci under study are given in Table S1. The frequency values observed in this study were compared with the previously published data (Table S2) on 17 autosomal STR markers from India and with recently published data on 22 autosomal STR markers .
The study will add to the data bank of various studies conducted on the Indian population. With respect to the distribution of alleles at each STR locus, the loci were found to be substantially polymorphic in this population indicating good informativeness of all studied autosomal STR markers. In addition, the Multiplex System is found suitable in deciphering the amelogenin deletion to avoid misinterpretation and/or to assess presence of male DNA (if the DNA is male mixed). The interested researchers can request the data from the authors.
The authors gratefully acknowledge the support of Director, State Forensic Science Laboratory, Sagar (MP) for providing laboratory facility and Promega (India) for providing the PowerPlex Fusion 6 C for testing its efficacy on the Indian population.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interest.
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