Mitochondrial 16S rRNA gene-dependent Blood typing as a Forensic Tool

Abstract

Background: Mitochondrial DNA is an important tool for human identification and is used to differentiate between human and animal blood at the crime scene, because in extreme conditions nuclear DNA is severely destroyed while Mitochondrial DNA contains multiple copies (200-2000) in per cell as well as resists harsh and more stable conditions. Methodology: Seventy-two blood samples were collected from human (Homo sapiens), sheep (Ovis aries), goat (Capra hircus) and cow (Bos taurus) (Eighteen blood samples for each). All blood samples were withdrawn by technician and 5ml were aspirated using aseptic technique and transferred to EDTA-Na2 tube and mixed well and stored in refrigerator. The collection takes 2 weeks (15th May 2019 to 30th May 2019). All samples were collected from Al-Diwanyia city. Results: The results of PCR reveal that, the primer pairs were specific and non-specific products not appear for all samples. The amplification of Homo sapiens mitochondrial DNA with primer pairs of other (Ovis aries, Capra hircus and Bos taurus) and amplification of each with primers pair of another genus gave negative results and this a primary evidence for primer pairs specificity. The amplicon of 16S rRNA gene of Homo sapiens were 1200bp ,Ovis aries were 1060bp ,Capra hircus were 820bp , and Bos taurus were 1300bp. The sequencing revealed that no crossreactivity of designed primer pairs and the PCR assay based on the designed primer pairs will be simple, fast, sensitive, specific, and cost-effective. Conclusion: Sensitivity, specificity and accuracy of the designed species specific primer pairs and applicability of the designed primer pairs in forensics to investigate blood sports or evidence belonging for human, sheep, goat and cow.