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80%

20%

0%

Positives out of N=41 Results

BLOOD %

SEMEN %

SALIVA %

0

10

20

30

40

50

60

POSITIVE RESULTS NA

50%

12,5%

21,9%

0

58,5%

0

10

20

Positive Negative

0

19

Phadebas 54.3 %

0

10

20

Positive Negative

0

14

RSIDTM-Saliva 40 %

0% 10% 20% 30% 40% 50% 60% 70% 80% 90%

Peroxidase

Immuno

RSIDTM-Blood (Glycophorin A)

OBTI® Hemoglobin

HemaTrace® Hemoglobin

RapidSignal Occult Blood-Hemoglobin

Hem-Direct® Hemoglobin

Hem-Check®

Teichmann crystals

HPLC

% Given results

Pruebas orientativas

• Induciendo el desprendimiento de oxígeno (capacidad del grupo Hemo para

la actividad peroxidasa). El O2 actúa en sustrato de bencidina (Adler), Luminol

(derivado ácido ftálico), Thevenon Roland-Piramidón

Pruebas de certeza

1.Cristalográficas (Teichmann, cristales de hematina; Tackayama: hemocromógeno)

2.Espectroscópicas (espectro absorción Hb y derivados)

3. Cromatográficas (TLC, HPLC)

Pruebas específicas Inmunológicas

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0

5

10

15

20

25

30

20

0 0 0 0 0 0 0

211

26

4

16

12 2

1

4

0

Positive

Negative

Inconclusive

Positive Negative Inconclusive

3

2 2

Alternate Lights 2 %

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4

0

5

10

15

20

25

0 0 0

4

10

23

4 4

Positive

Negative

Boward ES. 2015. Forensic Magazine. 05/21/2015. On the scene and In the lab.

10; 40%

2; 9%

11; 44%

2; 9%

Cytochrome b

sequencing

CO I,II sequencing

SPInDEL

STRs

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RecomendacionesLinacre et al. 2011. ISFG Recommendations

regarding the use of non-human (animal) DNA in

forensic genetic investigation. Forensic

Science International: Genetics. doi:10.1016/j.fsigen.2010.10.017

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6

92%

100%

50%

0%

20%

40%

60%

80%

100%

120%

Sequencing SPInDEL STRs

Positive identification N=24

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SPInDel multiplex

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Graphical representation of the six ribosomal RNA (rRNA)target regions in the mitochondrial DNA (mtDNA) amplifiedby multiplex PCR in the SPInDel profiling kit

Schematic illustration of the strategy used inthe species identification by theinsertions/deletions (SPInDel) method

Four conserved regions (green boxes) define threehypervariable domains (dotted brown lines). A section of thealignment is magnified to show the presence of multiple gapsin hypervariable regions. Each species is identified by anumeric profile resulting from the combination of lengths inhypervariable regions. Pereira F, Carneiro J, Matthiesen R, van Asch B, Pinto N, Gusmão L,

Amorim A. Identification of species by multiplex analysis of variable-length sequences. Nucleic Acids Res. 2010 Dec;38(22):e203.

The reference SPInDel profiles for the 10 target species are:

Species Markers SPID2716 SPID1350 SPID639 SPID1051 SPID2975 SPID2173 Capra hircus (Goat) 11 15 12 14 13 14 Canis lupus familiaris (Dog) 19 11 12 13 19 15 Equus caballus (Horse) 18 25 16 15 17 15 Oryctolagus cuniculus (Rabbit)17 11 12 13 16 17 Felis catus (Cat) 13 13 14 15 19 12 Homo sapiens (Human) 10 10 11 10 10 11 Ovis aries (Sheep) 11 15 13 14 12 16 Sus scrofa (Pig) 14 17 13 13 16 10 Mus musculus (Mouse) 12 10 11 15 15 14 Bos taurus (Cattle) 11 16 10 14 11 14

Carneiro J, Pereira F, Amorim A. Mol Ecol Resour. 2012 12(6):1190-5.

Pereira F, Carneiro J, Matthiesen R, van Asch B, Pinto N, Gusmão L, AmorimA. Nucleic Acids Res. 2010 Dec;38(22):e203.

Spanish and Portuguese-Speaking Working Group of the InternationalSociety for Forensic Genetics (GHEP-ISFG) http://www.gep-isfg.org/pt/comisses-trabalho/exercicio-colaborativo-ghep-isfg-8220spindel.html

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The reference SPInDel profiles for the species:

Species Markers SPID2716 SPID1350 SPID639 SPID1051 SPID2975 SPID2173 Sus scrofa (Pig) 14 17 13 13 16 10

Carneiro J, Pereira F, Amorim A. Mol Ecol Resour. 2012 12(6):1190-5.

Pereira F, Carneiro J, Matthiesen R, van Asch B, Pinto N, Gusmão L, AmorimA. Nucleic Acids Res. 2010 Dec;38(22):e203.

Spanish and Portuguese-Speaking Working Group of the InternationalSociety for Forensic Genetics (GHEP-ISFG) http://www.gep-isfg.org/pt/comisses-trabalho/exercicio-colaborativo-ghep-isfg-8220spindel.html

STRs identification: Descripción del número de alelos identificados en distintas poblaciones para cada marcador microsatélite

Marker Populations

1 2 3 4 5SW1370 5 6 10 7 10SW1824 6 5 8 6 10SW1891 7 3 6 5 10SW1904 5 6 8 8 12SW21 4 4 4 4 6SW2156 6 7 10 9 13SW2519 4 5 7 6 11SWR2527 5 5 9 7 10SW818 6 7 9 6 12SWR1829 9 6 9 9 12

Estudio intraespecífico. STRstetraméricos. Individualización.

Linacre et al. 2011. Forensic Science

International: Genetics. doi:10.1016/j.fsigen.2010.10.017

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Locus Fluorescent

labelSize

(bp)

Sequence of primers (5’-3’)*

SW24 FAM 19 F: CTT TGG GTG GAG TGT GTG CR: ATC CAA ATG CTG CAA GCG

SW936 HEX 21 F: TCT GGA GCT AGC ATA AGT GCCR: GTG CAA GTA CAC ATG CAG GG

S0355 HEX 26 F: TCT GGC TCC TAC ACT CCT TCT TGA TGR: GTT TGG GTG GGT GCT GAA AAA TAG GA

SW72 NED 18 F: ATC AGA ACA GTG CGC CGTR: TTT GAA AAT GGG GTG TTT CC

*Putnova et al. 2003. Czech Janim Sci., 48 (8): 307-314.

Lin et al. 2014. Forensic Sci. Int.: Genetics 9: 12-19Naue et al. 2014. Int J Legal Med. 128: 11-18

Rohrer et al. 2007. Animal Genetics 38: 253-258

Escena del crimen (especies domésticas)

Tráfico de especies exóticas

Dípteros (análisis PMI)

Fraude alimentario

Microbiología forense