title: Development of new PCR multiplex system by the simultaneous detection of 10 miniSTRs, SE33, Penta E, Penta D, and four Y-STRs.
abstract: The 18 loci multiplex system has been instigated for co-amplification and fluorescent detection of Amelogenin and 17 STRs, including 10 MiniSTRs (CSF1PO, D18S51, D7S820, D2S1338, TPOX, D13S317, FGA, D5S818, D21S11, D16S539), SE33, Penta E, Penta D, and four Y-STRs (DYS385a/b, DYS438, DYS392). This multiplex system was developed for the simultaneous analysis of compromised DNA samples, Y-amelogenin marker mutation, motherless paternity issues where single allele sharing occurs at autosomal STRs in unrelated individuals, and other complex forensic cases. Selection of loci, primers, and allelic ladders were designed and created in-house with a design strategy to work in this multiplex. The multiplex system was evaluated by sensitivity, specificity, stability, precision and accuracy, case-type samples, mixture studies, PCR-based and population distribution studies to establish the robustness and reliability of the system as the current requirements of the forensic case work. Among all the markers evaluated for this study, 209 alleles including 44 variants were observed with combined power of discrimination, combined power of exclusion, and the combined probability of matching calculated as 0.999999999999999999893916339344, 0.999993816173890, and 5.90019 × 10-19, respectively. Due to highly polymorphic characteristics of these loci particularly SE33 and Penta E which are most discriminatory (PD = 0.991 and 0.983, respectively) in the Pakistani population, this multiplex would be highly valuable for individual identification in complex forensic cases and paternity issues as well as population database.
authors.affiliation: Forensic DNA Typing Laboratory, National Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan. shafique@cemb.edu.pk.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shafique
authors.affiliation: Faculty of Life Sciences, University of Central Punjab, Lahore, Pakistan.
authors.forename: Muhammad Saqib
authors.initials: MS
authors.lastname: Shahzad
authors.affiliation: Forensic DNA Typing Laboratory, National Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Ziaur
authors.initials: Z
authors.lastname: Rahman
authors.affiliation: Forensic Services Laboratory, Centre for Applied Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Muhammad Adnan
authors.initials: MA
authors.lastname: Shan
authors.affiliation: Forensic Services Laboratory, Centre for Applied Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Rukhsana
authors.initials: R
authors.lastname: Perveen
authors.affiliation: Forensic Services Laboratory, Centre for Applied Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Shahzad
authors.affiliation: Forensic Services Laboratory, Centre for Applied Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Manzoor
authors.initials: M
authors.lastname: Hussain
authors.affiliation: Forensic DNA Typing Laboratory, National Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Ahmad Ali
authors.initials: AA
authors.lastname: Shahid
authors.affiliation: Forensic DNA Typing Laboratory, National Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, 53700, Pakistan.
authors.forename: Tayyab
authors.initials: T
authors.lastname: Husnain
mesh_terms: D053523:Amelogenin / Q000235:genetics; D000818:Animals; D041322:Chromosomes, Human, Y; D016172:DNA Fingerprinting; D005819:Genetic Markers; D005828:Genetics, Population; D005838:Genotype; D006801:Humans; D018598:Minisatellite Repeats; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013045:Species Specificity; D020080:Tandem Repeat Sequences
chemical_list: D053523:Amelogenin; D005819:Genetic Markers
keywords: Forensic; MiniSTRs; PCR multiplex; Penta D; Penta E; SE33; Y-STRs