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1. RNA/DNA co-analysis from human skin and contact traces – results of a sixth collaborative EDNAP exercise

Author

M. van den Berge, A.M. Bento, Angel Carracedo, Christine Keyser, S.A. Harbison, Katja Drobnič, Titia Sijen, Maria João Porto, Eva Sauer, Marielle Vennemann, Peter M. Schneider, Erin K. Hanson, C. Franchi, Niels Morling, Jack Ballantyne, Gavrilo Hadzic, Clémence Hollard, G. De Cock, R. Fleming, Fabrice Noel, P. Miniati, Cordula Haas, Olalla Maroñas, Walther Parson, Andrea Berti, Cornelius Courts, Harald Niederstätter, G. Shanthan, M. Turanská, Athina Vidaki, L. Zatkalíková, D. Syndercombe Court, Nicola A. McCallum, P. Hoff-Olsen, Benjamin Benn Hjort, A.D. Roeder, R. Banemann, A. Kondili, and Iva Gomes

Subjects
Forensic Genetics, Messenger RNA, RNA, Human skin, DNA, Biology, Molecular biology, Pathology and Forensic Medicine, Housekeeping gene, chemistry.chemical_compound, DNA profiling, chemistry, Reference genes, Genetics, Humans, RNA extraction, Skin
Abstract

The European DNA profiling group (EDNAP) organized a sixth collaborative exercise on RNA/DNA co-analysis for body fluid/tissue identification and STR profiling. The task was to identify skin samples/contact traces using specific RNA biomarkers and test three housekeeping genes for their suitability as reference genes. Eight stains, a skin RNA dilution series and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by 22 participating laboratories using RNA extraction or RNA/DNA co-extraction methods. Two sets of previously described skin-specific markers were used: skin1 pentaplex (LCE1C, LCE1D, LCE2D, IL1F7 and CCL27) and skin2 triplex (LOR, KRT9 and CDSN) in conjunction with a housekeeping gene, HKG, triplex (B2M, UBC and UCE). The laboratories used different chemistries and instrumentation. All laboratories were able to successfully isolate and detect mRNA in contact traces (e.g., human skin, palm-, hand- and fingerprints, clothing, car interiors, computer accessories and electronic devices). The simultaneous extraction of RNA and DNA provides an opportunity for positive identification of the tissue source of origin by mRNA profiling as well as a simultaneous identification of the body fluid donor by STR profiling. The skin markers LCE1C and LOR and the housekeeping gene marker B2M were detected in the majority of contact traces. Detection of the other markers was inconsistent, possibly due to the low amounts and/or poor quality of the genetic material present in shed skin cells. The results of this and the previous collaborative RNA exercises support RNA profiling as a reliable body fluid/tissue identification method that can easily be combined with current STR typing technology.

Published
2015
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2. RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: results of a fourth and fifth collaborative EDNAP exercise

Author

C. Franchi, Eva Sauer, M. Dötsch, Katja Drobnič, Mónica Carvalho, Peter M. Schneider, A.D. Roeder, Niels Morling, Olalla Maroñas, Clémence Hollard, Christine Keyser, R.A.H. van Oorschot, Iva Gomes, S.A. Harbison, R. Fleming, M. Turanská, Marielle Vennemann, Gavrilo Hadzic, Christopher Phillips, Erin K. Hanson, R. Banemann, Athina Vidaki, Kaye N. Ballantyne, D. Moore, L. Zatkalíková, Walther Parson, Benjamin Benn Hjort, Joyce Harteveld, Fabrice Noel, L. Salvaderi, Harald Niederstätter, Cornelius Courts, M.J. Anjos, G. Shanthan, Cordula Haas, C. Popielarz, Jack Ballantyne, G. De Cock, D. Syndercombe Court, Nicola A. McCallum, P. Hoff-Olsen, Eduardo Euclydes de Lima e Borges, C. Hüls, and Bryan Bhoelai

Subjects
Body fluid, Messenger RNA, RNA, DNA, Biology, Molecular biology, Pathology and Forensic Medicine, Housekeeping gene, Body Fluids, Menstruation, chemistry.chemical_compound, Blood, DNA profiling, chemistry, Reference genes, Vagina, Genetics, Humans, Female, RNA extraction
Abstract

The European DNA Profiling Group (EDNAP) organized a fourth and fifth collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling. The task was to identify dried menstrual blood and vaginal secretion stains using specific RNA biomarkers, and additionally test 3 housekeeping genes for their suitability as reference genes. Six menstrual blood and six vaginal secretion stains, two dilution series (1/4-1/64 pieces of a menstrual blood/vaginal swab) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by 24 participating laboratories, using RNA extraction or RNA/DNA co-extraction methods. Two novel menstrual blood mRNA multiplexes were used: MMP triplex (MMP7, MMP10, MMP11) and MB triplex (MSX1, LEFTY2, SFRP4) in conjunction with a housekeeping gene triplex (B2M, UBC, UCE). Two novel mRNA multiplexes and a HBD1 singleplex were used for the identification of vaginal secretion: Vag triplex (MYOZ1, CYP2B7P1 and MUC4) and a Lactobacillus-specific Lacto triplex (Ljen, Lcris, Lgas). The laboratories used different chemistries and instrumentation and all were able to successfully isolate and detect mRNA in dried stains. The simultaneous extraction of RNA and DNA allowed for positive identification of the tissue/fluid source of origin by mRNA profiling as well as a simultaneous identification of the body fluid donor by STR profiling, also from old and compromised casework samples. The results of this and the previous collaborative RNA exercises support RNA profiling as a reliable body fluid identification method that can easily be combined with current STR typing technology.

Published
2013
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3. A global analysis of Y-chromosomal haplotype diversity for 23 STR loci

Author

Thomas Capal, Marian M. de Pancorbo, Birgit Bayer, Sheila M.T. Angustia, Francesca Brisighelli, Andrea Piccinini, Lutz Roewer, Yali Xue, Aikaterini Kondili, Alexandra Ampati, Katja Anslinger, Mouayyad Al-Azem, Witold Pepinski, Harald Niederstätter, Jae Joseph Russell B. Rodriguez, Katja Drobnič, Denise Syndercombe Court, Jana Wobst, David Ballard, Danel Rey-González, Marina Nastainczyk-Wulf, Christopher Phillips, Magdalena Konarzewska, Evelyn K. Guevara, Jill K. Olofsson, Maria Eugenia D’Amato, Natsuko Mizuno, Ansar El Andari, Yiping Hou, G.M. Luque, Carolyn R. Hill, Antti Sajantila, Cordula Haas, Tobi Gail Smith, Christian Gehrig, Andreas O. Tillmar, Marilidia Piglionica, Anja E. Klann, M. Sirker, Jazelyn M. Salvador, Jukka U. Palo, Rita Y.Y. Yong, Antonio Alonso, Xinia Barrantes, Bruce Budowle, Helen Burri, Xianping Wang, Baowen Cheng, Paulina Wolańska-Nowak, Peter de Knijff, Renato Salazar, Thirsa Kraaijenbrink, Shenglan Chen, Susi Pelotti, Tatiana Lúcia Santos Nogueira, Mark A. Jobling, Iulia Skitsa, Issam Mansour, Jeanett Edelmann, Urs V. Borer, Stefania Lonero Baldassarra, Maria Corazon A. De Ungria, Di Zhou, William E. Frank, Jon H. Wetton, Begoña Martínez-Jarreta, Sheng-Ping Hu, Marion Nagy, Andrea Verzeletti, Rodrigo Soares de Moura Neto, Martin R. Whittle, Anna Jonkisz, Ryszard Pawłowski, Cristian Capelli, Yu Na Oh, Emila Jastrzebska, Helena Nilsson, Agnieszka Maciejewska, M. Kohl, Cristina Cano García, Miguel Marino, Michael D. Coble, Elizeu Fagundes de Carvalho, Gareth M. Gwynne, Michael Nothnagel, Chengtao Jiang, Tadeusz Dobosz, Gavrilo Hadzic, Burkhard Berger, Sascha Willuweit, Branka Grskovic, Katsuya Honda, Tanja Ilievska, Rüdiger Lessig, Zlatko Jakovski, Antonio Salas, Balázs Egyed, Laura Locarno, Christoph Koller, Helena Moreira, Sibylle Tschumi, Carolina Núñez, Luís Souto Miranda, Nicola Schlauderer, Patrícia Domingues, Yasuki Iwashima, Di Lu, V. Cortellini, Antónia Völgyi, Monica Abreu-Głowacka, Mariela Caputo, Maarten Larmuseau, Peter M. Schneider, Wafaa Takash Chamoun, Gerhard Bäßler, Pekka Saukko, Cíntia Alves, Lejla Kovacevic, Maria Vouropoulou, Walther Parson, Stefania Turrina, Chris Tyler-Smith, Lina Solis De Calvit, D.R. Sumita, Qasim Ayub, T. Wiest, Iris Lindner, Jorge Cárdenas, Kyoung Jin Shin, Daniel Corach, Claudio Ottoni, Carlo Robino, Sabine Siegert, Ivana Furač, Miriam Baeta Bafalluy, Uta Dorothee Immel, Domenico De Leo, Ronny Decorte, Sandra Furfuro, Niels Morling, Valerio Onofri, Adriano Tagliabracci, Rosane Silva, Beate Balitzki, Ulises Toscanini, Olga Rickards, Michael Krawczak, Sean Davison, Ulrike Schmidt, Wei Wei, Pablo Martín, Vincent Castella, Yan Li, P. Miniati, Rafał Płoski, Gusztáv Bárány, Vlastimil Stenzl, Shengjie Nie, Horolma Pamjav, Carey Davis, Josephine Purps, Lorna H. Santos, Damir Marjanović, Hjelt Institute (-2014), Forensic Medicine, PaleOmics Laboratory, Purps, Josephine, Siegert, Sabine, Willuweit, Sascha, Nagy, Marion, Alves, Cíntia, Salazar, Renato, Angustia, Sheila M.T., Santos, Lorna H., Anslinger, Katja, Bayer, Birgit, Ayub, Qasim, Wei, Wei, Xue, Yali, Tyler-Smith, Chri, Bafalluy, Miriam Baeta, Martínez-Jarreta, Begoña, Egyed, Balaz, Balitzki, Beate, Tschumi, Sibylle, Ballard, David, Court, Denise Syndercombe, Barrantes, Xinia, Bäßler, Gerhard, Wiest, Tina, Berger, Burkhard, Niederstätter, Harald, Parson, Walther, Davis, Carey, Budowle, Bruce, Burri, Helen, Borer, Ur, Koller, Christoph, Carvalho, Elizeu F., Domingues, Patricia M., Chamoun, Wafaa Takash, Coble, Michael D., Hill, Carolyn R., Corach, Daniel, Caputo, Mariela, D'Amato, Maria E., Davison, Sean, Decorte, Ronny, Larmuseau, Maarten H.D., Ottoni, Claudio, Rickards, Olga, Lu, Di, Jiang, Chengtao, Dobosz, Tadeusz, Jonkisz, Anna, Frank, William E., Furac, Ivana, Gehrig, Christian, Castella, Vincent, Grskovic, Branka, Haas, Cordula, Wobst, Jana, Hadzic, Gavrilo, Drobnic, Katja, Honda, Katsuya, Hou, Yiping, Zhou, Di, Li, Yan, Hu, Shengping, Chen, Shenglan, Immel, Uta-Dorothee, Lessig, Rüdiger, Jakovski, Zlatko, Ilievska, Tanja, Klann, Anja E., García, Cristina Cano, De Knijff, Peter, Kraaijenbrink, Thirsa, Kondili, Aikaterini, Miniati, Penelope, Vouropoulou, Maria, Kovacevic, Lejla, Marjanovic, Damir, Lindner, Iri, Mansour, Issam, Al-Azem, Mouayyad, Andari, Ansar El, Marino, Miguel, Furfuro, Sandra, Locarno, Laura, Martín, Pablo, Luque, Gracia M., Alonso, Antonio, Miranda, Luís Souto, Moreira, Helena, Mizuno, Natsuko, Iwashima, Yasuki, Neto, Rodrigo S. Moura, Nogueira, Tatiana L.S., Silva, Rosane, Nastainczyk-Wulf, Marina, Edelmann, Jeanett, Kohl, Michael, Nie, Shengjie, Wang, Xianping, Cheng, Baowen, Núñez, Carolina, Pancorbo, Marian Martínez De, Olofsson, Jill K., Morling, Niel, Onofri, Valerio, Tagliabracci, Adriano, Pamjav, Horolma, Volgyi, Antonia, Barany, Gusztav, Pawlowski, Ryszard, Maciejewska, Agnieszka, Pelotti, Susi, Pepinski, Witold, Abreu-Glowacka, Monica, Phillips, Christopher, Cárdenas, Jorge, Rey-Gonzalez, Danel, Salas, Antonio, Brisighelli, Francesca, Capelli, Cristian, Toscanini, Ulise, Piccinini, Andrea, Piglionica, Marilidia, Baldassarra, Stefania L., Ploski, Rafal, Konarzewska, Magdalena, Jastrzebska, Emila, Robino, Carlo, Sajantila, Antti, Palo, Jukka U., Guevara, Evelyn, Salvador, Jazelyn, Ungria, Maria Corazon De, Rodriguez, Jae Joseph Russell, Schmidt, Ulrike, Schlauderer, Nicola, Saukko, Pekka, Schneider, Peter M., Sirker, Miriam, Shin, Kyoung-Jin, Oh, Yu Na, Skitsa, Iulia, Ampati, Alexandra, Smith, Tobi-Gail, Calvit, Lina Solis De, Stenzl, Vlastimil, Capal, Thoma, Tillmar, Andrea, Nilsson, Helena, Turrina, Stefania, De Leo, Domenico, Verzeletti, Andrea, Cortellini, Venusia, Wetton, Jon H., Gwynne, Gareth M., Jobling, Mark A., Whittle, Martin R., Sumita, Denilce R., Wolańska-Nowak, Paulina, Yong, Rita Y.Y., Krawczak, Michael, Nothnagel, Michael, and Roewer, Lutz

Subjects
Forensic Genetics, Population structure, RECOMBINATION, Forensic Genetic, purl.org/becyt/ford/1 [https], AMOVA, Database, Discriminatory power, Gene diversity, Population structure, Genética y Herencia, 0302 clinical medicine, MARKERS, Haplotype, POPULATION, Allele, Genetics, 0303 health sciences, education.field_of_study, Medicine (all), Gene diversity, 319 Forensic science and other medical sciences, 16. Peace & justice, Gene diversity, Discriminatory power, AMOVA, Population structure, Database, Str loci, Microsatellite Repeat, AMOVA, Database, Discriminatory power, Gene diversity, Population structure, Genetics, Forensic Medicine, ALLELE FREQUENCIES, Discriminatory power, CIENCIAS NATURALES Y EXACTAS, Human, AMOVA, Population, Settore BIO/08 - ANTROPOLOGIA, Biology, Settore BIO/08, SEQUENCE, Article, Pathology and Forensic Medicine, EVENTS, Ciencias Biológicas, Database, 03 medical and health sciences, Genetic, Settore MED/43 - Medicina Legale, Humans, 030216 legal & forensic medicine, education, purl.org/becyt/ford/1.6 [https], Allele frequency, Alleles, 030304 developmental biology, Chromosomes, Human, Y, ta1184, DELETION, Null (mathematics), AZFA REGION, ta3121, Haplotypes, 3111 Biomedicine, SYSTEM, Microsatellite Repeats
Abstract

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent. Fil: Corach, Daniel. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Servicio de Huellas Digitales Genéticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; 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; Argentina Fil: Marino, Miguel Eduardo. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Analisis de ADN; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Purps, Josephine. Charité-Universitätsmedizin; Alemania Fil: Siegert, Sabine. University of Cologne; Alemania Fil: Willuweit, Sascha. Charité-Universitätsmedizin; Alemania Fil: Nagy, Marion. Charité-Universitätsmedizin; Alemania Fil: Alves, Cíntia. Universidad de Porto; Portugal Fil: Salazar, Renato. Universidad de Porto; Portugal Fil: Angustia, Sheila M. T.. Philippine National Police Crime Laboratory; Filipinas Fil: Santos, Lorna H.. Philippine National Police Crime Laboratory; Filipinas Fil: Anslinger, Katja. Universitat Genzentrum Der Ludwing-maximilians; Alemania Fil: Bayer, Birgit. Universitat Genzentrum Der Ludwing-maximilians; Alemania Fil: Ayub, Qasim. The Wellcome Trust Sanger Institute; Reino Unido Fil: Wei, Wei. The Wellcome Trust Sanger Institute; Reino Unido Fil: Xue, Yali. The Wellcome Trust Sanger Institute; Reino Unido Fil: Tyler Smith, Chris. The Wellcome Trust Sanger Institute; Reino Unido Fil: Baeta Bafalluy, Miriam. Universidad de Zaragoza; España Fil: Martínez Jarreta, Begoña. Universidad de Zaragoza; España Fil: Egyed, Balazs. Eotvos University, Budapest; Argentina Fil: Balitzki, Beate. Universidad de Basilea; Suiza Fil: Tschumi, Sibylle. Universidad de Basilea; Suiza Fil: Ballard, David. King; Reino Unido Fil: Syndercombe Court, Denise. King; Reino Unido Fil: Barrantes, Xinia. Poder Judicial, Forensic Sciences Department; Costa Rica Fil: Bäßler, Gerhard. Landeskriminalamt Baden-Württemberg; Alemania Fil: Berger, Burkhard. Universidad de Innsbruck; Austria Fil: Niederstätter, Haral. Universidad de Innsbruck; Austria Fil: Parson, Walther. Universidad de Innsbruck; Austria. University Park; Estados Unidos Fil: Davis, Carey. Department of Molecular and Medical Genetics; Estados Unidos. Institute of Applied Genetics; Estados Unidos Fil: Furfuro, Sandra. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentina Fil: Locarno, Laura. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Laboratorio de Análisis de ADN; Argentina

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