Lee, Ji Eun, Lee, Jeong Min, Naue, Jana, Fleckhaus, Jan, Freire-Aradas, Ana ORCID: 0000-0002-8135-5182, Neubauer, Jacqueline ORCID: 0000-0002-9472-7803, Pospiech, Ewelina, McCord, Bruce, Kalamara, Vivian, Gauthier, Quentin, Mills, Carly, Cao, Yijian, Wang, Zheng, Oh, Yu Na, Feng, Lei, Schneider, Peter M. ORCID: 0000-0003-0744-2349, Phillips, Christopher, Haas, Cordula ORCID: 0000-0001-8122-1427, Pisarek, Aleksandra, Branicki, Wojciech, Podini, Daniele, Vidaki, Athina ORCID: 0000-0002-5470-245X, Tejero, Nicole Fernandez, Ambroa-Conde, Adrian, Mosquera-Miguel, Ana, Lareu, Maria Victoria, Hou, Yiping, Lee, Joo Young and Lee, Hwan Young (2022). A collaborative exercise on DNA methylation-based age prediction and body fluid typing. Forensic Sci. Int.-Genet., 57. CLARE: ELSEVIER IRELAND LTD. ISSN 1878-0326

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Abstract

DNA methylation has become one of the most useful biomarkers for age prediction and body fluid identification in the forensic field. Therefore, several assays have been developed to detect age-associated and body fluid-specific DNA methylation changes. Among the many methods developed, SNaPshot-based assays should be particularly useful in forensic laboratories, as they permit multiplex analysis and use the same capillary electrophoresis instrumentation as STR analysis. However, technical validation of any developed assays is crucial for their proper integration into routine forensic workflow. In the present collaborative exercise, two SNaPshot multiplex assays for age prediction and a SNaPshot multiplex for body fluid identification were tested in twelve laboratories. The experimental set-up of the exercise was designed to reflect the entire workflow of SNaPshot-based methylation analysis and involved four increasingly complex tasks designed to detect potential factors influencing methylation measurements. The results of body fluid identification from each laboratory provided sufficient information to determine appropriate age prediction methods in subsequent analysis. In age prediction, systematic measurement differences resulting from the type of genetic analyzer used were identified as the biggest cause of DNA methylation variation between laboratories. Also, the use of a buffer that ensures a high ratio of specific to non-specific primer binding resulted in changes in DNA methylation measurement, especially when using degenerate primers in the PCR reaction. In addition, high input volumes of bisulfite-converted DNA often caused PCR failure, presumably due to carry-over of PCR inhibitors from the bisulfite conversion reaction. The proficiency of the analysts and experimental conditions for efficient SNaPshot reactions were also important for consistent DNA methylation measurement. Several bisulfite conversion kits were used for this study, but differences resulting from the use of any specific kit were not clearly discerned. Even when different experimental settings were used in each laboratory, a positive outcome of the study was a mean absolute age prediction error amongst participant's data of only 2.7 years for semen, 5.0 years for blood and 3.8 years for saliva.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Lee, Ji EunUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Lee, Jeong MinUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Naue, JanaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Fleckhaus, JanUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Freire-Aradas, AnaUNSPECIFIEDorcid.org/0000-0002-8135-5182UNSPECIFIED
Neubauer, JacquelineUNSPECIFIEDorcid.org/0000-0002-9472-7803UNSPECIFIED
Pospiech, EwelinaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
McCord, BruceUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Kalamara, VivianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Gauthier, QuentinUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mills, CarlyUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Cao, YijianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wang, ZhengUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Oh, Yu NaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Feng, LeiUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schneider, Peter M.UNSPECIFIEDorcid.org/0000-0003-0744-2349UNSPECIFIED
Phillips, ChristopherUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Haas, CordulaUNSPECIFIEDorcid.org/0000-0001-8122-1427UNSPECIFIED
Pisarek, AleksandraUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Branicki, WojciechUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Podini, DanieleUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Vidaki, AthinaUNSPECIFIEDorcid.org/0000-0002-5470-245XUNSPECIFIED
Tejero, Nicole FernandezUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ambroa-Conde, AdrianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mosquera-Miguel, AnaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Lareu, Maria VictoriaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hou, YipingUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Lee, Joo YoungUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Lee, Hwan YoungUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-693483
DOI: 10.1016/j.fsigen.2021.102656
Journal or Publication Title: Forensic Sci. Int.-Genet.
Volume: 57
Date: 2022
Publisher: ELSEVIER IRELAND LTD
Place of Publication: CLARE
ISSN: 1878-0326
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
IDENTIFICATION; MARKERS; SEMEN; PATTERNS; SALIVAMultiple languages
Genetics & Heredity; Medicine, LegalMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/69348

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