Developing Applications for Forensic Epigenetics using Pyrosequencing, Real-Time PCR and Other Methods Involving Laboratory Based Genetic Analysis

Bruce McCord, Analytical and Forensic Chemistry, Florida International University, Florida , USA (lili@bioforum.co.il)

We have developed a set of epigenetic markers which produce unique and specific patterns of DNA methylation that can be used to identify blood, semen, saliva, sweat, and vaginal epithelial cells. These new methods easily fit within the workflow of a standard forensic DNA laboratory. Our procedure involves testing DNA extracted from human samples, which is then amplified using bisulfite modified PCR. Specific primers amplify the region of interest, and the quantitative methylation profile of each locus is determined by pyrosequencing. The versatility of these new markers is presented by showing the results of validation studies on sensitivity, human specificity, stability, and mixture resolution. We also will demonstrate recent results on the specificity of saliva markers. Saliva, composed of different cell types, exhibits differences in methylation percent depending on the collection method. Thus, a buccal/lip swab is different than spit, potentially allowing the differentiation between deposition methods. Lastly, we present additional data on the utilization of pyrosequencing for age and other phenotypic information such as smoking status. The results of these methods can provide important information in criminal casework.

Short Biography of Presenting Author

Bruce R. McCord is a Professor of Analytical and Forensic Chemistry at Florida International University. Dr. McCord received a BS in Chemistry with honors from the College of William and Mary in 1981, and a Ph.D. in Analytical Chemistry from the University of Wisconsin-Madison in 1986. Prior to working at FIU, he was an Assistant Professor at Ohio University (1998-2004) and a Research Chemist at the FBI Laboratory (1989-1998).

His current research interests involve the development of procedures for forensic analysis including both biochemical (genomics) and chemical assays (trace analysis). This work includes projects in forensic epigenetics, rapid PCR, pyrosequecing, microfluidics and surface enhanced Raman spectroscopy. Dr. McCord has published over 130 peer reviewed papers, 12 patents, 16 book chapters and has graduated 31 PhD students.

He serves as Deputy Editor for the journal Electrophoresis, Editor for Forensic Sciences (MDPI) and is a member of the Editorial Board of the Journal of Forensic Chemistry as well as the scientific committee of the Green Mountain DNA Conference. In 2008 he received the Paul Kirk Award from the American Academy of Forensic Sciences. When not in the laboratory he can be found performing improvisational jazz on the saxophone and clarinet or foil windsurfing in the Florida Keys.

17-18 JANUARY 2023, THE DAVID INTERCONTINENTAL HOTEL, TEL AVIV, ISRAEL
Developing Applications for Forensic Epigenetics using Pyrosequencing, Real-Time PCR and Other Methods Involving Laboratory Based Genetic Analysis Bruce McCord, Analytical and Forensic Chemistry, Florida International University, Florida , USA (lili@bioforum.co.il) We have developed a set of epigenetic markers which produce unique and specific patterns of DNA methylation that can be used to identify blood, semen, saliva, sweat, and vaginal epithelial cells. These new methods easily fit within the workflow of a standard forensic DNA laboratory. Our procedure involves testing DNA extracted from human samples, which is then amplified using bisulfite modified PCR. Specific primers amplify the region of interest, and the quantitative methylation profile of each locus is determined by pyrosequencing. The versatility of these new markers is presented by showing the results of validation studies on sensitivity, human specificity, stability, and mixture resolution. We also will demonstrate recent results on the specificity of saliva markers. Saliva, composed of different cell types, exhibits differences in methylation percent depending on the collection method. Thus, a buccal/lip swab is different than spit, potentially allowing the differentiation between deposition methods. Lastly, we present additional data on the utilization of pyrosequencing for age and other phenotypic information such as smoking status. The results of these methods can provide important information in criminal casework. Short Biography of Presenting Author Bruce R. McCord is a Professor of Analytical and Forensic Chemistry at Florida International University. Dr. McCord received a BS in Chemistry with honors from the College of William and Mary in 1981, and a Ph.D. in Analytical Chemistry from the University of Wisconsin-Madison in 1986. Prior to working at FIU, he was an Assistant Professor at Ohio University (1998-2004) and a Research Chemist at the FBI Laboratory (1989-1998). His current research interests involve the development of procedures for forensic analysis including both biochemical (genomics) and chemical assays (trace analysis). This work includes projects in forensic epigenetics, rapid PCR, pyrosequecing, microfluidics and surface enhanced Raman spectroscopy. Dr. McCord has published over 130 peer reviewed papers, 12 patents, 16 book chapters and has graduated 31 PhD students. He serves as Deputy Editor for the journal Electrophoresis, Editor for Forensic Sciences (MDPI) and is a member of the Editorial Board of the Journal of Forensic Chemistry as well as the scientific committee of the Green Mountain DNA Conference. In 2008 he received the Paul Kirk Award from the American Academy of Forensic Sciences. When not in the laboratory he can be found performing improvisational jazz on the saxophone and clarinet or foil windsurfing in the Florida Keys.

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