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TinLizzy
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Date Posted:12/31/2010 12:47 AMCopy HTML

The Portuguese Laboratory appeared to have analysed quite a number of samples. This is just one post from a 9 page thread on themaddiecasefiled.com (translations by Albym, I understand):

NATIONAL INSTITUTE OF LEGAL MEDICINE
GENETIC AND BIOLOGICAL FORENSIC SERVICES

Processo n° 2007/000268/P-B
Processo n° 2007/000244/CR-B
Processo n° 2007/000226/LX-BC

EXPERT REPORT

IDENTIFICATION OF BIOLOGICAL TRACES AND COMPARATIVE STUDY WITH REFERENCE SAMPLES

INFORMATION


REQUESTING AGENCY: Departamento de Investigacao Criminal de Portimao da Policia Judiciaria e Directoria Nacional.

PROCESSO: NUIPC 201/07.0GALGS
NOME: MADELEINE BETH MCCANN.

OFICIO N.° 013.079 and fax n° 399, both dated 08/05/2007, that say "to proceed to deliver the traces recovered from apartment n° 5-A of the tourist resort "The Ocean Club" in Praia da Luz in Lagos", which were accompanied by 7 (seven) sealed envelopes under a delivery note n° 156/2007-CR/L, as part of the investigation into the disappearance of the child MADELEINE BETH MCCANN.

OFICIO N.° 13443 and fax n° 413 both dated 15/05/2007, that say to proceed to deliver the traces recovered from the residence at Rua do Ramalhete, Casa Liliana in Praia da Luz in Lagos (1 envelope), and from the Volkswagen with numberplate 44-77-KD (7 sealed envelopes), which were accompanied by delivery notes n° 170A/2007CR/L and 170B/2007-CR/L. respectively.

FAX N° 000415 dated 15/05/2007, that requests the INML to proceed with all necessary examinations on all the traces delivered to them as part of the abovementioned Inquiry.

OFICIO N.° 13453 /I /2007 dated 15/05/2007, that says to proceed to deliver the bucal swabs recovered from Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick Mccann, Kate Marie Healy, with delivery note n° 170C/2007-CR/L and relevant official documents from the Departamento de Investigacao Criminal de Portimao da Policia Judiciaria.

OFICIO N.° 001803 dated 17/05/07, that says "to remit bucal swabs recovered from individuals, asking for comparisons of DNA profiles with those from biological traces recovered from various locations, as part of the present legal documents", accompanied by relevant official documents from the Departamento de lnvestigacao Criminal de Portimao da Policia Judiciaria.

OFICIO N.° 002026 dated 05/06/07, that says "to deliver two bucal swabs provided by Robert James Queriol Eveleigh Murat", with relevant official documents from the Departamento de Investigacao Criminal de Portimao da Policia Judiciaria.

MATERIAL RECEIVED


There were received 258 traces and 12 reference samples - bucal swabs from 11 individuals.

DESCRIPTION OF MATERIAL RECEIVED


TRACES
Relating to Apartment n° 5-A in the tourist resort "The Ocean Club" Praia da Luz-Lagos:

- 32 hairs in envelope n° 1 recovered from the floor at the entrance to the children's bedroom.

- 28 hairs in envelope n° 2 recovered from the floor next to the bed from which the child disappeared.

- 4 hair in envelope n° 3 recovered from the top of the bed from which the child disappeared.

- 15 hairs in envelope n° 4 recovered from the floor next to the bed that was next to the window in the children's bedroom.

- 1 piece of cloth in envelope n° 5 recovered from the bedspread of the bed next to the window in the children's bedroom.
Fragment of cloth, mauve/violet in colour with square motifs, circular in form about 10cm in diameter. A small fluorescent spot is observed under a Crime-light.

- 31 hairs in envelope N° 6 recovered from the floor of the lounge.

- 58 hairs in envelope N° 7 recovered from the entrance hall at the front door of the apartment.

[The above were] Delivered by the Policia Judiciaria on 08/05/2007.

Relating to the Volkswagen 44-77-KD:

- 15 hairs in envelope N° 1 recovered from the front-right seat.

- 2 hairs in envelope N° 2 recovered from the front-left seat.

- 12 hairs in N° 3 recovered from the floor in front of the front seats.

- 7 hairs in envelope N° 4 recovered from the floor in front of the rear seat.

- 4 hairs in envelope N° 5 recovered from the rear seat.

- 5 hairs in envelope N° 6 recovered from the rear part of the vehicle.

- 4 hairs in envelope N° 7 recovered from the internal covering of the roof.

Relating to Residencia Casa Liliana - Praia da Luz-Lagos:

- 40 hairs in envelope N° 1 recovered from the bed in the bedroom of of the suspect.

[The above were] Delivered by the Policia Judiciaria on 15/05/2007.

REFERENCE SAMPLES

- 4 sealed bucal swabs from:
Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick Mccann, Kate Marie Healey.

Delivered by the Policia Judiciaria on 15/05/2007.

- 6 sealed bucal swabs from:
Dianne Webster, Rachael Mariamma Jean Mampilly, Russell James O'Brien, Fiona Payne, David Anthony Payne, Silvia Maria Correia Batista.

Delivered by the Policia Judiciaria on 18/05/2007.

- 2 sealed bucal swabs from:
Robert James Queriol Eveleigh Murat.

Remitted by the Departamento de Investigacao Criminal de Portimao da Policia Judiciaria and delivered on 06/06/2007.

METHODOLOGY

1-Morphology analysis of hairs

- Macro- and microscopic characterisation.

2-Nature of the sample

Acid Phosphatase Test to detect semen on the small spot on the cloth fragment in envelope n° 5 recovered from the bedspread of the bed next to the window of the children's bedroom.
[Ref: http://www.semenonpanties.com/laux.htm ]

3- DNA extraction:

The following methods were used to extract DNA from the material received for analysis:

- Extraction with phenol-chloroform (organic extraction) [1,2]
[Ref: http://en.wikipedia.org/wiki/Phenol-chl ... extraction ]
- Tissue and Hair Extraction kit -"DNA IQ(tm) System" (Promega)
- "QIAamp micro DNA kit" method (Qiagen)
- Chelex method [3]

[1] Huhne, J., Pfiffer, H., Waterkamp, K., Brinkmann, B. 1999. "Mitochondrial DNA in human hair shafts existence of intra-individual differences?". International Journal of Legal Medicine (1999), 112: 172-175.
[2] Maniatis T. Fritsch EF, Sambrook J. 1982. "Molecular Cloning - A Laboratory Manual". Cold Spring Harbor Laboratory.
[3] Walsh P.S., Metzer D.A., Higuchi (1991). Chelex-100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10: 506-513.

4-Nuclear DNA study

After extraction the following genetic markers were studied by PCR:

· Autosomic STRs: D8S1179, D21S11, D7S820, CSFlPO, D3S1358, HUMTH01, D13S317, D16S539, D2S1338, D19S433, HUMVWA31/A, TPOX, D18S51, D5S818, HUMFIBRA/FGA and Amelogene, using the primers and conditions of amplification suggested in the kit AmpFISTR Identifiler (Applied Bioystems).

· Autosomic STRs: D3S1358, HUMTH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSFlPO, Penta D, HUMVWA31/A, D8S1179, TPOX, HUMFIBRA/FGA, and Amelogene using the primers and the conditions of amplification suggested in kit PowerPlex(tm)16 System (Promega).

5-Mitochondrial DNA study

The following methodology was used in the Mitochondrial DNA study:

PCR Amplification:

PCR Amplification of the human mtDNA regions HVI and HVII, using specified primers and amplification conditions in accordance with Vigilant et al., 1989 [4] e Wilson et al., 1995 [5], Alvarez-lglesias et al., [6].

[4] Vigilant L, Pennington R. Harpending 11, Kocher TD, Wilson AC. 1989. "Mitochondrial DNA Sequences in Single Hairs from a Southern African Population". Proc Natl Acad Sci, USA, 1989, Vol. 86: 9350 - 9354.
[5] Wilson MR, DiZinno JA, Polanskey D, Replogle J. Budowle B. 1995. "Validation of Mitochondrial DNA Sequencing for Forensic Casework Analysis". Int J Legal Med. 1995,108: 68 - 74.
[6] Alvarez-Iglesias V, Jaime JC, Carracedo A, Salas A.2007. "Coding region mitochondrial DNA SNPs:Targeting East Asian and Native American haplogroups" Forensic Sci. Int. Genetics, 2007, 1 :44-55.

Sequenciation:
Direct sequenciation of the HVI e HVll segments by capilar electrophoresis in "ABI PRISM 3100 ou 3130 Genetic Analyser" (Applied Biosystems) automated sequenciator with fluorescent detection using the following kits:

- "BigDye(R) Terminator vl.1 Cycle Sequencing" (Applied Biosystems)
- "BigDye(R) Terminator v1.3 Cycle Sequencing" (Applied Biosystems)
- "dRhodamine Terminator Cycle Sequencing Ready Reaction" (Applied Biosystems)

The programme "SeqScape(R) Software Version 2.0" (Applied Biosystems) was used for comparison of the sequences obtained from the analysed samples with the CRS (Cambridge Reference Sequence).

RESULTS


1- Morphology analysis of the hairs

All hairs received for analysis were subjected to macro- and microscopic characterisation, with the majority of those having the root being in a telogenic phase.
[Ref: https://www.promega.com/geneticidproc/e ... c/ab20.pdf ]

Relating to Apartment n° 5-A

- Envelope N° 1 - "recovered from the floor at the entrance to the children's bedroom" 25 hairs with root; 4 hairs only stem; 3 hairs non-human.

- Envelope N° 2 - "recovered from the floor next to the bed from which the child disappeared" 3 hairs with root; 25 hairs only stem.

- Envelope N° 3 - "recovered from the top of the bed from which the child disappeared" 2 hairs with root; 2 hairs only stem.

- Envelope N° 4 - " recovered from the floor next to the bed that was next to the window in the children's bedroom" ll hairs with root; 4 hairs only stem.

- Envelope n° 6 - "recovered from the floor of the lounge" 26 hairs with root; 4 hairs only stem; l hair non-human.

- Envelope n° 7 - "recovered from the entrance hall at the front door of the apartment" 44 hairs with root; 8 hairs only stem; 6 hairs non-human.

Relating to the Volkswagen 44-77-KD

- Envelope n° 1 - "recovered from the front-right seat" 13 hairs with root; 1 hair root only; 1 hair non-human.

- Envelope n° 2 - "recovered from the front-left seat" 2 hairs with root.

- Envelope n° 3 - "recovered from the floor in front of the front seats" 9 hairs with root; 2 hairs only stem; 1 hair non-human.

- Envelope N° 4 - "recovered from the floor in front of the rear seat" 1 hair with root; 6 hairs only stem.

- Envelope N° 5 - "recovered from the rear seat" 4 hairs only stem.

- Envelope N° 6 - "recovered from the rear part of the vehicle" 1 hair with root; 4 hairs only stem.

- Envelope N° 7 - "recovered from the internal covering of the roof" 3 hairs with root; 1 hair only stem.

Relating to the Residencia Casa Liliana - Praia da Luz-Lagos:

- Envelope N° 1 - "recovered from the bed in the bedroom of of the suspect" 36 hairs with root; 4 hairs only stem.

2-Nature of the sample

Acid Phosphatase Test to detect semen on the small spot on the cloth fragment in envelope n° 5 recovered from the bedspread of the bed next to the window of the children's bedroom: Weakly positive.

3 - Nuclear DNA study

Autosomic STRs:

- were obtained from: hair root CEnv 7-51 recovered in the apartment (entrance hall-Env.7), hair root PVT07 recovered in the vehicle (floor in front of rear seat Env.4), hair root C742-05 recovered in the vehicle (floor in front of the front seat- Env. 3) and the small spot on the piece of cloth (recovered from the bedspread of the bed next to the window in the children's bedroom Env.5).

- were obtained from swabs:

Matthew David Oldfield, Jane Michelle Tanner, Gerald Patrick Mccann, Kate Marie Healey, Dianne Webster, Rachael Mariamma Jean Mampilly, Russell James O'Brien, Fiona Payne, David Anthony Payne, Silvia Maria Correia Batista e Robert James Queriol Eveleigh Murat.

4 - Mitochondrial DNA study

- Mitochondrial DNA was obtained from 194 vestiges - 1 spot and 193 hairs (roots and/or stems) of the 246 analysed. It was not possible to obtain results from 52 hairs, 31 of which were recovered in the apartment.


Mitochondrial DNA profiles of the vestiges
Code:
mtDNA Number of Comparison with
profile Samples Recovered from reference samples

B 3 3 entrance hall
B1 1 1 entrance hall
-------------------------------------------------------
13 lounge
15 entrance hall
C 53 13 entrance to bedroom Haplotype of Kate Healy (Mother)
4 floor; bed next to window
1 bed of the child
7 floor; next to child's bed
-------------------------------------------------------
D 1 1 lounge
D1 1 1 entrance to bedroom
D1 1 1 floor; bed next to window
-------------------------------------------------------
F 3 1 lounge
2 entrance hall
-------------------------------------------------------
G 1 1 lounge Haplotipo de Matthew Oldfield
I 1 1 entrance hall Haplotipo de David Payne
-------------------------------------------------------
1 floor; bed next to window
J 3 1 entrance hall
1 floor; next to child's bed
-------------------------------------------------------
K 14 1 Residencia Liliana
13 vehicle(envlps 1,3,4,5 and 6)
-------------------------------------------------------
1 spot on bedspread
L 8 1 entrance to bedroom
2 entrance hall
4 floor; bed next to window
-------------------------------------------------------
M 48 13 vehicle (envlps 1,2,3 and 4)
35 Residencia Liliana Haplotipo de Robert Murat
-------------------------------------------------------
2 lounge
11 entrance hall
N 24 6 entrance to bedroom Haplotipo de Gerald Mccann
2 floor; bed next to window
3 floor; next to child's bed
-------------------------------------------------------
0 2 1 lounge
1 entrance hall Haplotipo de Russell O'Brien
-------------------------------------------------------
Q 2 1 entrance hall
1 lounge
-------------------------------------------------------
Profiles different from each other and from those above:
28 17 of which were found in the apartment.

* Bl and Dl have a different sequence from B and D.

CONCLUSIONS


1st- The macro- and microscopic analysis of 257 hairs revealed 245 human and 12 non-humano.

2nd- With respect to autosomic STRs the genetic profiles of the following analysed samples matched with the reference sample indicated::

- hair root CEnv 7-51 recovered in the apartment (entrance hall-Env.7) matched with Gerald Mccann, father of the victim.

- hair root PVT07 recovered in the vehicle (floor in front of rear seat Env.4) match with Robert James Queriol Eveleigh Murat.

3rd- With respect to autosomic STRs the female genetic profile identified from hair root C742-05 recovered in the vehicle(floor in front of the front seat- Env 3) did not match any profile from the reference samples.

4th- With respect to autosomic STRs the male genetic profile identified from the spot on the cloth fragment (bedspread of the bed next to window in children's bedroom- Env 5) did not match any profile from the reference samples.

5th- In the samples from apartment 5-A, several mitochondrial DNA profiles were found:

- Profile identified by letter "C", present in 53 samples, was identical to that of Kate Healy, mother of the victim, meaning those samples were from her or from someone having the same maternal bloodline.

- Profile identified by letter "N", present in 24 samples, was identical to that of Gerald Mccann, father of the victim, meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "G", present in 1 samples, was identical to that of Matthew David Oldfield, meaning that sample was from him or from someone having the same maternal bloodline.

- Profile identified by letter "I", present in 1 samples, was identical to that of David Anthony Payne, meaning that sample was from him or from someone having the same maternal bloodline.

- Profile identified by letter "O", present in 2 samples, was identical to that of Russell James O'Brien, meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "L", present in the spot on the bedspread of the bed next to the window and in seven hairs, meaning that all these samples came from the same person or from someone having the same maternal bloodline, did not match any of the reference samples.

- Profiles identified by letters "B", "D", "F", "J" and "Q" are different from the above, and from each other, and are distinct from reference samples.

6th- In the samples analysed from Residencia Liliana and the vehicle, several mitochondrial DNA profiles were found:

- Profile identified by letter "M", present in 48 samples (35 from Residencia Liliana and 13 from the vehicle), was identical to that of Robert James Queriol Eveleigh Murat meaning those samples were from him or from someone having the same maternal bloodline.

- Profile identified by letter "K", present in 14 samples (1 from Residencia Liliana and 13 from the vehicle), meaning that all these samples came from the same person or from someone having the same maternal bloodline, did not match any of the reference samples.

- One of the 13 samples from the vehicle in letter "K" - hair root C742-05 (floor in front of the front seat - Env 3) showed a female DNA genetic profile.

7th- The remaining 28 samples analysed, of which 17 were recovered from the apartment, showed mitochondrial DNA from different from each other, and distinct from those above.

8th- Of the 245 hairs analysed, no results were obtained from 52.

National Institute of Legal Medicine, 9 July 2007.

[Titles, names and signatures of signatories to the report]


http://themaddiecasefiles.com/topic171.html?hilit=forensic%20lab

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