Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise

S Ingold; G Dørum; E Hanson; A Berti; W Branicki; P Brito; P Elsmore; K B Gettings; F Giangasparo; T E Gross; S Hansen; E N Hanssen; M-L Kampmann; M Kayser; F-X Laurent; N Morling; A Mosquera-Miguel; W Parson; C Phillips; M J Porto; E Pośpiech; A D Roeder; P M Schneider; K Schulze Johann; C R Steffen; D Syndercombe-Court; M Trautmann; M van den Berge; K J van der Gaag; J Vannier; V Verdoliva; A Vidaki; C Xavier; J Ballantyne; C Haas

doi:10.1016/j.fsigen.2018.01.002

Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise

S Ingold, G Dørum, E Hanson, A Berti, W Branicki, P Brito, P Elsmore, K B Gettings, F Giangasparo, T E Gross, S Hansen, E N Hanssen, M-L Kampmann, M Kayser, F-X Laurent, N Morling, A Mosquera-Miguel, W Parson, C Phillips, M J PortoE Pośpiech, A D Roeder, P M Schneider, K Schulze Johann, C R Steffen, D Syndercombe-Court, M Trautmann, M van den Berge, K J van der Gaag, J Vannier, V Verdoliva, A Vidaki, C Xavier, J Ballantyne, C Haas

Section of Forensic Genetics

26 Citations (Scopus)

Abstract

In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.

Original language	English
Journal	Forensic science international. Genetics
Volume	34
Pages (from-to)	105-115
Number of pages	11
ISSN	1872-4973
DOIs	https://doi.org/10.1016/j.fsigen.2018.01.002
Publication status	Published - May 2018

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Ingold, S., Dørum, G., Hanson, E., Berti, A., Branicki, W., Brito, P., Elsmore, P., Gettings, K. B., Giangasparo, F., Gross, T. E., Hansen, S., Hanssen, E. N., Kampmann, M.-L., Kayser, M., Laurent, F.-X., Morling, N., Mosquera-Miguel, A., Parson, W., Phillips, C., ... Haas, C. (2018). Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise. Forensic science international. Genetics, 34, 105-115. https://doi.org/10.1016/j.fsigen.2018.01.002

Ingold, S, Dørum, G, Hanson, E, Berti, A, Branicki, W, Brito, P, Elsmore, P, Gettings, KB, Giangasparo, F, Gross, TE, Hansen, S, Hanssen, EN, Kampmann, M-L, Kayser, M, Laurent, F-X, Morling, N, Mosquera-Miguel, A, Parson, W, Phillips, C, Porto, MJ, Pośpiech, E, Roeder, AD, Schneider, PM, Schulze Johann, K, Steffen, CR, Syndercombe-Court, D, Trautmann, M, van den Berge, M, van der Gaag, KJ, Vannier, J, Verdoliva, V, Vidaki, A, Xavier, C, Ballantyne, J & Haas, C 2018, 'Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise', Forensic science international. Genetics, vol. 34, pp. 105-115. https://doi.org/10.1016/j.fsigen.2018.01.002

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title = "Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise",

abstract = "In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.",

author = "S Ingold and G D{\o}rum and E Hanson and A Berti and W Branicki and P Brito and P Elsmore and Gettings, {K B} and F Giangasparo and Gross, {T E} and S Hansen and Hanssen, {E N} and M-L Kampmann and M Kayser and F-X Laurent and N Morling and A Mosquera-Miguel and W Parson and C Phillips and Porto, {M J} and E Po{\'s}piech and Roeder, {A D} and Schneider, {P M} and {Schulze Johann}, K and Steffen, {C R} and D Syndercombe-Court and M Trautmann and {van den Berge}, M and {van der Gaag}, {K J} and J Vannier and V Verdoliva and A Vidaki and C Xavier and J Ballantyne and C Haas",

year = "2018",

month = may,

doi = "10.1016/j.fsigen.2018.01.002",

language = "English",

volume = "34",

pages = "105--115",

journal = "Forensic science international. Genetics",

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T1 - Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise

AU - Ingold, S

AU - Dørum, G

AU - Hanson, E

AU - Berti, A

AU - Branicki, W

AU - Brito, P

AU - Elsmore, P

AU - Gettings, K B

AU - Giangasparo, F

AU - Gross, T E

AU - Hansen, S

AU - Hanssen, E N

AU - Kampmann, M-L

AU - Kayser, M

AU - Laurent, F-X

AU - Morling, N

AU - Mosquera-Miguel, A

AU - Parson, W

AU - Phillips, C

AU - Porto, M J

AU - Pośpiech, E

AU - Roeder, A D

AU - Schneider, P M

AU - Schulze Johann, K

AU - Steffen, C R

AU - Syndercombe-Court, D

AU - Trautmann, M

AU - van den Berge, M

AU - van der Gaag, K J

AU - Vannier, J

AU - Verdoliva, V

AU - Vidaki, A

AU - Xavier, C

AU - Ballantyne, J

AU - Haas, C

PY - 2018/5

Y1 - 2018/5

N2 - In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.

AB - In a previous study we presented an assay for targeted mRNA sequencing for the identification of human body fluids, optimised for the Illumina MiSeq/FGx MPS platform. This assay, together with an additional in-house designed assay for the Ion Torrent PGM/S5 platform, was the basis for a collaborative exercise within 17 EUROFORGEN and EDNAP laboratories, in order to test the efficacy of targeted mRNA sequencing to identify body fluids. The task was to analyse the supplied dried body fluid stains and, optionally, participants' own bona fide or mock casework samples of human origin, according to specified protocols. The provided primer pools for the Illumina MiSeq/FGx and the Ion Torrent PGM/S5 platforms included 33 and 29 body fluid specific targets, respectively, to identify blood, saliva, semen, vaginal secretion, menstrual blood and skin. The results demonstrated moderate to high count values in the body fluid or tissue of interest with little to no counts in non-target body fluids. There was some inter-laboratory variability in read counts, but overall the results of the laboratories were comparable in that highly expressed markers showed high read counts and less expressed markers showed lower counts. We performed a partial least squares (PLS) analysis on the data, where blood, menstrual blood, saliva and semen markers and samples clustered well. The results of this collaborative mRNA massively parallel sequencing (MPS) exercise support targeted mRNA sequencing as a reliable body fluid identification method that could be added to the repertoire of forensic MPS panels.

U2 - 10.1016/j.fsigen.2018.01.002

DO - 10.1016/j.fsigen.2018.01.002

M3 - Journal article

C2 - 29453107

SN - 1872-4973

VL - 34

SP - 105

EP - 115

JO - Forensic science international. Genetics

JF - Forensic science international. Genetics

ER -

Body fluid identification using a targeted mRNA massively parallel sequencing approach - results of a EUROFORGEN/EDNAP collaborative exercise

Abstract

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