ABO and Amelogenin Determination by PCR, from Experimental Bloodstains and from Museum Specimens, Using a Non-Destructive Approach

Abstract

Background/Objectives: Museum collections constitute valuable material for investigating a wide range of histological processes. This results from the historical selection of unusual and advanced disease cases by museum curators, which are rarely encountered in contemporary clinical practice due to advances in medicine. Ancient DNA plays a crucial role in phylogenetic studies, as well as in analyses of population genetics. However, many commonly used DNA extraction techniques involve partial degradation of samples prior to DNA isolation. The use of non-destructive methods may enable the recovery of DNA appropriate for downstream analyses. Non-destructive methods of DNA extraction for research purposes are a recent development and facilitate genetic analyses of museum collections. ABO and Amel are examples of applications of the proposed method, although any set of primers can be used. ABO genotyping has been widely used in phylogenetic and population analyses. Methods: This study presents a non-destructive approach for PCR-based DNA extraction from preserved museum samples. Human tissue samples, filter materials used during preservation, and processed conservation fluids (after dilution and dialysis) were analyzed to determine ABO genotype and sex (based on Amelogenin). Results: The same replicable PCR profile (ABO blood group and sex determined by Amelogenin) was observed across all three sample types: tissues, filter papers, and conservation fluid. The use of a preservative solution is a new development, as it leaves the sample intact. However, this approach has a drawback: DNA diffuses into the preservative solution very slowly, and it takes several decades to reach a sufficient concentration. A major advantage of this approach is the ability to perform a PCR test without DNA preparation. Conclusions: Museum-derived samples represent a reliable source of DNA and can be effectively used in PCR-based analyses. The presented method works well with degraded DNA samples, combining an already established very short Amelogenin amplicon with PCR sequence-specific primers for ABO genotyping. Keywords: museum specimens, ABO typing, PCR, multiplex, LOH, MSI