Species identification in forensic samples using the SPInDel approach: A GHEP-ISFG inter-laboratory collaborative exercise

• Autores: Alves, Cíntia; Pereira, Rui; Prieto, Lourdes; Aler, Mercedes; Amaral, Cesar R. L.; Arévalo, Cristina; Berardi, Gabriela; Di Rocco, Florencia; Caputo, Mariela; Carmona, Cristian Hernandez; Catelli, Laura; Costa, Heloísa Afonso; Coufalova, Pavla; Furfuro, Sandra Beatriz; García, Óscar; Gaviria, Anibal; Goios, Ana; Gómez, Juan José Builes; Hernández, Alexis; Betancor Hernández, Eva del Carmen; Miranda, Luís; Parra, David; Pedrosa, Susana; Porto, Maria João Anjos; Rebelo, Maria de Lurdes; Spirito, Matteo; Torres, María del Carmen Villalobos; Amorim, António; Pereira, Filipe
• Año de publicación: 2017
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: Collaborative Exercise; Forensic Investigations; Mtdna; Species Identification; Spindel
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

DNA is a powerful tool available for forensic investigations requiring identification of species. However, it is necessary to develop and validate methods able to produce results in degraded and or low quality DNA samples with the high standards obligatory in forensic research. Here, we describe a voluntary collaborative exercise to test the recently developed Species Identification by Insertions/Deletions (SPInDel) method. The SPInDel kit allows the identification of species by the generation of numeric profiles combining the lengths of six mitochondrial ribosomal RNA (rRNA) gene regions amplified in a single reaction followed by capillary electrophoresis. The exercise was organized during 2014 by a Working Commission of the Spanish and Portuguese-Speaking Working Group of the International Society for Forensic Genetics (GHEP-ISFG), created in 2013. The 24 participating laboratories from 10 countries were asked to identify the species in 11 DNA samples from previous GHEP-ISFG proficiency tests using a SPInDel primer mix and control samples of the 10 target species. A computer software was also provided to the participants to assist the analyses of the results. All samples were correctly identified by 22 of the 24 laboratories, including samples with low amounts of DNA (hair shafts) and mixtures of saliva and blood. Correct species identifications were obtained in 238 of the 241 (98.8%) reported SPInDel profiles. Two laboratories were responsible for the three cases of misclassifications. The SPInDel was efficient in the identification of species in mixtures considering that only a single laboratory failed to detect a mixture in one sample. This result suggests that SPInDel is a valid method for mixture analyses without the need for DNA sequencing, with the advantage of identifying more than one species in a single reaction. The low frequency of wrong (5.0%) and missing (2.1%) alleles did not interfere with the correct species identification, which demonstrated the advantage of using a method based on the analysis of multiple loci. Overall, the SPInDel method was easily implemented by laboratories using different genotyping platforms, the interpretation of results was straightforward and the SPInDel software was used without any problems. The results of this collaborative exercise indicate that the SPInDel method can be applied successfully in forensic casework investigations.
Fil: Alves, Cíntia. Universidad de Porto; Portugal
Fil: Pereira, Rui. Universidad de Porto; Portugal
Fil: Prieto, Lourdes. Universidad de Santiago de Compostela; España
Fil: Aler, Mercedes. Instituto de Medicina Legal y Ciencias Forenses de Valencia; España
Fil: Amaral, Cesar R. L.. Universidade do Estado do Rio de Janeiro; Brasil
Fil: Arévalo, Cristina. Universidad de Alcalá; España
Fil: Berardi, Gabriela. Fundación Favaloro; Argentina
Fil: Di Rocco, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentina
Fil: Caputo, Mariela. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Carmona, Cristian Hernandez. Poder Judicial. Departamento de Ciencias Forenses. Sección de Bioquímica; Costa Rica
Fil: Catelli, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Equipo Argentino de Antropología Forense; Argentina
Fil: Costa, Heloísa Afonso. Instituto Nacional de Medicina Legal e Ciências Forenses; Portugal
Fil: Coufalova, Pavla. Institute of Criminalistics Prague; República Checa
Fil: Furfuro, Sandra Beatriz. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentina
Fil: García, Óscar. Polícia del País Vasco. Sección de Genética Forense; España
Fil: Gaviria, Anibal. Cruz Roja Ecuatoriana; Ecuador
Fil: Goios, Ana. Universidad de Porto; Portugal
Fil: Gómez, Juan José Builes. Universidad de Antioquia; Colombia
Fil: Hernández, Alexis. Instituto Nacional de Toxicología y Ciencias Forenses; España
Fil: Betancor Hernández, Eva del Carmen. Instituto de Medicina Legal de Las Palmas. Laboratorio Genética Forense; España
Fil: Miranda, Luís. Universidade de Aveiro; Portugal
Fil: Parra, David. Servicio de Criminalística de la Guardia Civil. Departamento de Química y Medio Ambiente; España
Fil: Pedrosa, Susana. Unidad de Laboratorio de Navarra de Servicios y Tecnologías; España
Fil: Porto, Maria João Anjos. Instituto Nacional de Medicina Legal e Ciências Forenses; Portugal
Fil: Rebelo, Maria de Lurdes. Instituto Nacional de Medicina Legal e Ciências Forenses; Portugal
Fil: Spirito, Matteo. Università Cattolica del Sacro Cuore; Italia
Fil: Torres, María del Carmen Villalobos. Universidad Autónoma de Nuevo León; México
Fil: Amorim, António. Universidad de Porto; Portugal
Fil: Pereira, Filipe. Universidad de Porto; Portugal