The genetic difference between male and female is determined by the presence of a Y chromosome in males, so the detection of specific sequences on this chromosome allows us to distinguish between male and female individuals. With the development of the polymerase chain reaction (PCR), DNA analysis to determine sex can be performed using small amounts of DNA in the forensic field and on various biological artifacts, including bones and archaeological remains. Polymerase chain reaction (PCR) is a fast and inexpensive procedure that allows for the amplification of single target DNA molecules in analytical quantities. The advent of molecular genetics has allowed for the development of more sensitive methods to determine sex. However, anthropometric studies provide good results only when the skeletons are complete, in good condition and of adult individuals. So far, determining sex of human remains has been based on the sexual dimorphism present in most human bones. Sex determination of archaeological human remains is of fundamental importance for anthropological studies of ancient societies, cultures and genealogical histories. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: This research received Institutional funding to Anna Poma, Alfonso Forgione, Antonella Bonfigli, Patrizia Cesare from University of L’Aquila.The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors declare no conflict of interest. Received: OctoAccepted: Published: June 10, 2022Ĭopyright: © 2022 Poma et al. PLoS ONE 17(6):Įditor: David Caramelli, University of Florence, ITALY (2022) A qPCR-duplex assay for sex determination in ancient DNA. The proposed molecular technique proves to be sensitive and precise even on degraded DNA, in fact on 9 archaeological finds dating from the VII-XII century in which sex had been identified through anthropometric analysis, it confirmed the sex of 8 out of 9 finds correctly.Ĭitation: Poma A, Cesare P, Bonfigli A, Volpe AR, Colafarina S, Vecchiotti G, et al.
The validity of the method was verified on modern DNA in which gender was identified in all the samples with 100% accuracy thus, allowing for the same results as the classic method with amelogenin, but in a faster and more immediate way, as it allows for sex determination solely by analyzing the denaturation curves without having to perform an electrophoretic run. This method involves the co-amplification of two genes in a single reaction system and the subsequent analysis of the fusion curves the gene sequences used for the construction of suitable primers are those of steroid sulfatase (STS) and testis specific protein Y-linked 1 (TSPY) genes which turned out to be two sensitive markers as they have a detection limit of 60 pg and 20 pg respectively on modern DNA.
In the present work, we investigated the possibility of determining sex by using the qPCR-duplex method for both ancient and modern DNA samples. Molecular biology techniques are increasingly being used in sex identification of skeletal remains when traditional anthropometric analyzes are not successful in identifying sex of remains that are incomplete, fragmented and /or of immature individuals.
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