Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group

Autores
Montesino, M.; Sala, Adriana Andrea; Crespillo, M.; Albarrán, C.; Alonso, A.; Álvarez Iglesias, V.; Cano, J. A.; Carvalho, M.; Corach, Daniel; Cruz, C.; Di Lonardo, A.; Espinheira, R.; Farfán, M. J.; Filippini, S.; García Hirschfeld, J.; Hernández, A.; Lima, G.; López Cubría, C. M.; López Soto, M.; Pagano, S.; Paredes, M.; Pinheiro, M. F.; RodríguezMonge, A. M.; Sala, A.; Sóñora, S.; Sumita, D. R.; Vide, M. C.; Whittle, M. R.; Zurita, A.; Prieto, L.
Año de publicación
2007
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.
Fil: Montesino, M.. No especifíca;
Fil: Sala, Adriana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Crespillo, M.. Instituto Nacional de Toxicologia y Ciencias Forenses; España
Fil: Albarrán, C.. Instituto Nacional de Toxicologia y Ciencias Forenses; España
Fil: Alonso, A.. Universidad de Santiago de Compostela; España
Fil: Álvarez Iglesias, V.. No especifíca;
Fil: Cano, J. A.. No especifíca;
Fil: Carvalho, M.. No especifíca;
Fil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cruz, C.. No especifíca;
Fil: Di Lonardo, A.. No especifíca;
Fil: Espinheira, R.. No especifíca;
Fil: Farfán, M. J.. No especifíca;
Fil: Filippini, S.. No especifíca;
Fil: García Hirschfeld, J.. No especifíca;
Fil: Hernández, A.. No especifíca;
Fil: Lima, G.. No especifíca;
Fil: López Cubría, C. M.. No especifíca;
Fil: López Soto, M.. No especifíca;
Fil: Pagano, S.. No especifíca;
Fil: Paredes, M.. No especifíca;
Fil: Pinheiro, M. F.. No especifíca;
Fil: RodríguezMonge, A. M.. No especifíca;
Fil: Sala, A.. No especifíca;
Fil: Sóñora, S.. No especifíca;
Fil: Sumita, D. R.. No especifíca;
Fil: Vide, M. C.. No especifíca;
Fil: Whittle, M. R.. No especifíca;
Fil: Zurita, A.. No especifíca;
Fil: Prieto, L.. No especifíca;
Materia
HAPLOTYPE
HVS-I
MTDNA MIXTURE
MTDNA PHYLOGENY
PREFERENTIAL LYSIS
SEQUENCING
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/162437

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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working groupMontesino, M.Sala, Adriana AndreaCrespillo, M.Albarrán, C.Alonso, A.Álvarez Iglesias, V.Cano, J. A.Carvalho, M.Corach, DanielCruz, C.Di Lonardo, A.Espinheira, R.Farfán, M. J.Filippini, S.García Hirschfeld, J.Hernández, A.Lima, G.López Cubría, C. M.López Soto, M.Pagano, S.Paredes, M.Pinheiro, M. F.RodríguezMonge, A. M.Sala, A.Sóñora, S.Sumita, D. R.Vide, M. C.Whittle, M. R.Zurita, A.Prieto, L.HAPLOTYPEHVS-IMTDNA MIXTUREMTDNA PHYLOGENYPREFERENTIAL LYSISSEQUENCINGhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.Fil: Montesino, M.. No especifíca;Fil: Sala, Adriana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Crespillo, M.. Instituto Nacional de Toxicologia y Ciencias Forenses; EspañaFil: Albarrán, C.. Instituto Nacional de Toxicologia y Ciencias Forenses; EspañaFil: Alonso, A.. Universidad de Santiago de Compostela; EspañaFil: Álvarez Iglesias, V.. No especifíca;Fil: Cano, J. A.. No especifíca;Fil: Carvalho, M.. No especifíca;Fil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cruz, C.. No especifíca;Fil: Di Lonardo, A.. No especifíca;Fil: Espinheira, R.. No especifíca;Fil: Farfán, M. J.. No especifíca;Fil: Filippini, S.. No especifíca;Fil: García Hirschfeld, J.. No especifíca;Fil: Hernández, A.. No especifíca;Fil: Lima, G.. No especifíca;Fil: López Cubría, C. M.. No especifíca;Fil: López Soto, M.. No especifíca;Fil: Pagano, S.. No especifíca;Fil: Paredes, M.. No especifíca;Fil: Pinheiro, M. F.. No especifíca;Fil: RodríguezMonge, A. M.. No especifíca;Fil: Sala, A.. No especifíca;Fil: Sóñora, S.. No especifíca;Fil: Sumita, D. R.. No especifíca;Fil: Vide, M. C.. No especifíca;Fil: Whittle, M. R.. No especifíca;Fil: Zurita, A.. No especifíca;Fil: Prieto, L.. No especifíca;Elsevier Ireland2007-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/162437Montesino, M.; Sala, Adriana Andrea; Crespillo, M.; Albarrán, C.; Alonso, A.; et al.; Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group; Elsevier Ireland; Forensic Science International; 168; 1; 5-2007; 42-560379-0738CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S037907380600449Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.forsciint.2006.06.066info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:06:53Zoai:ri.conicet.gov.ar:11336/162437instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:06:53.889CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
title Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
spellingShingle Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
Montesino, M.
HAPLOTYPE
HVS-I
MTDNA MIXTURE
MTDNA PHYLOGENY
PREFERENTIAL LYSIS
SEQUENCING
title_short Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
title_full Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
title_fullStr Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
title_full_unstemmed Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
title_sort Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group
dc.creator.none.fl_str_mv Montesino, M.
Sala, Adriana Andrea
Crespillo, M.
Albarrán, C.
Alonso, A.
Álvarez Iglesias, V.
Cano, J. A.
Carvalho, M.
Corach, Daniel
Cruz, C.
Di Lonardo, A.
Espinheira, R.
Farfán, M. J.
Filippini, S.
García Hirschfeld, J.
Hernández, A.
Lima, G.
López Cubría, C. M.
López Soto, M.
Pagano, S.
Paredes, M.
Pinheiro, M. F.
RodríguezMonge, A. M.
Sala, A.
Sóñora, S.
Sumita, D. R.
Vide, M. C.
Whittle, M. R.
Zurita, A.
Prieto, L.
author Montesino, M.
author_facet Montesino, M.
Sala, Adriana Andrea
Crespillo, M.
Albarrán, C.
Alonso, A.
Álvarez Iglesias, V.
Cano, J. A.
Carvalho, M.
Corach, Daniel
Cruz, C.
Di Lonardo, A.
Espinheira, R.
Farfán, M. J.
Filippini, S.
García Hirschfeld, J.
Hernández, A.
Lima, G.
López Cubría, C. M.
López Soto, M.
Pagano, S.
Paredes, M.
Pinheiro, M. F.
RodríguezMonge, A. M.
Sala, A.
Sóñora, S.
Sumita, D. R.
Vide, M. C.
Whittle, M. R.
Zurita, A.
Prieto, L.
author_role author
author2 Sala, Adriana Andrea
Crespillo, M.
Albarrán, C.
Alonso, A.
Álvarez Iglesias, V.
Cano, J. A.
Carvalho, M.
Corach, Daniel
Cruz, C.
Di Lonardo, A.
Espinheira, R.
Farfán, M. J.
Filippini, S.
García Hirschfeld, J.
Hernández, A.
Lima, G.
López Cubría, C. M.
López Soto, M.
Pagano, S.
Paredes, M.
Pinheiro, M. F.
RodríguezMonge, A. M.
Sala, A.
Sóñora, S.
Sumita, D. R.
Vide, M. C.
Whittle, M. R.
Zurita, A.
Prieto, L.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv HAPLOTYPE
HVS-I
MTDNA MIXTURE
MTDNA PHYLOGENY
PREFERENTIAL LYSIS
SEQUENCING
topic HAPLOTYPE
HVS-I
MTDNA MIXTURE
MTDNA PHYLOGENY
PREFERENTIAL LYSIS
SEQUENCING
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.
Fil: Montesino, M.. No especifíca;
Fil: Sala, Adriana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Crespillo, M.. Instituto Nacional de Toxicologia y Ciencias Forenses; España
Fil: Albarrán, C.. Instituto Nacional de Toxicologia y Ciencias Forenses; España
Fil: Alonso, A.. Universidad de Santiago de Compostela; España
Fil: Álvarez Iglesias, V.. No especifíca;
Fil: Cano, J. A.. No especifíca;
Fil: Carvalho, M.. No especifíca;
Fil: Corach, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cruz, C.. No especifíca;
Fil: Di Lonardo, A.. No especifíca;
Fil: Espinheira, R.. No especifíca;
Fil: Farfán, M. J.. No especifíca;
Fil: Filippini, S.. No especifíca;
Fil: García Hirschfeld, J.. No especifíca;
Fil: Hernández, A.. No especifíca;
Fil: Lima, G.. No especifíca;
Fil: López Cubría, C. M.. No especifíca;
Fil: López Soto, M.. No especifíca;
Fil: Pagano, S.. No especifíca;
Fil: Paredes, M.. No especifíca;
Fil: Pinheiro, M. F.. No especifíca;
Fil: RodríguezMonge, A. M.. No especifíca;
Fil: Sala, A.. No especifíca;
Fil: Sóñora, S.. No especifíca;
Fil: Sumita, D. R.. No especifíca;
Fil: Vide, M. C.. No especifíca;
Fil: Whittle, M. R.. No especifíca;
Fil: Zurita, A.. No especifíca;
Fil: Prieto, L.. No especifíca;
description The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.
publishDate 2007
dc.date.none.fl_str_mv 2007-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/162437
Montesino, M.; Sala, Adriana Andrea; Crespillo, M.; Albarrán, C.; Alonso, A.; et al.; Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group; Elsevier Ireland; Forensic Science International; 168; 1; 5-2007; 42-56
0379-0738
CONICET Digital
CONICET
url http://hdl.handle.net/11336/162437
identifier_str_mv Montesino, M.; Sala, Adriana Andrea; Crespillo, M.; Albarrán, C.; Alonso, A.; et al.; Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group; Elsevier Ireland; Forensic Science International; 168; 1; 5-2007; 42-56
0379-0738
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S037907380600449X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.forsciint.2006.06.066
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Ireland
publisher.none.fl_str_mv Elsevier Ireland
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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