1. Lutz Roewer, Dept. Forensic Genetics
The golden age of forensic Y-STR typing – new markers, new databases, new technologies
Lutz Roewer, Dept. Forensic Genetics
Institute of Legal Medicine and Forensic Sciences, Charité – Universitätsmedizin Berlin
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

2. PowerPlex Y (12 Loci) Promega Y-Filer (17 Loci) Applied Biosystems
Minimal Haplotype
PowerPlex Y (12 Loci) Promega
Y-Filer (17 Loci) Applied Biosystems
PPY23 (23 Loci) Promega
YFiler plus beta (27 Loci) Applied Biosystems
Commercial +
add. Y-STRs (45 loci)
DYS19 x
DYS389I
DYS389II
DYS390
DYS391
DYS392
DYS393
DYS385ab (2)
DYS437
DYS438
DYS439
DYS448
DYS456
DYS458
DYS635
YGATAH4
DYS570
DYS576
DYS533
DYS481
DYS549 x 
DYS643  x
DYS460
DYS627
DYS518
DYS449
DYF387S1ab (2)
DYF399S1 (3)
DYF403S1 (3)
DYF404S1 (2)
DYS526
DYS547
DYS612
DYS626
DYS464 (4)
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

3. Really fast mutating Y-STR markers are very rare
Ballantyne KN, Goedbloed M, Fang R, Schaap O, Lao O, Wollstein A, Choi Y, van Duijn K, Vermeulen M, Brauer S, Decorte R, Poetsch M, von Wurmb-Schwark N, de Knijff P, Labuda D, Vézina H, Knoblauch H, Lessig R, Roewer L, Ploski R, Dobosz T, Henke L, Henke J, Furtado MR, Kayser M (2010) Mutability of Y-chromosomal microsatellites: rates, characteristics, molecular bases, and forensic implications. Am J Hum Genet 87(3):
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

4. Family analysed with 45 markers – patrilineage resolved with 2 mutations
576 3 8 9 I 4 II 1
481
549
533
438
437
570
635
390
439
392
643
393
458 5 ab 6 G A T H 2 7
460
518
449 S
526a
526b
626 S1
612
547 F
DYS391 µ=2.54 x 10-3
DYS626 µ=1.22 x 10-2
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

5. Yfiler® Plus 25 marker, 2 bilocal 6 dye matrix
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

6. Ranking of mutation rates (Yfiler® Plus)
Loci
Mutation Rate [95% CI]
Meioses
Position[MutRate]
Group[MutRate]
dys438
2,96E-04
10122 1
slow
dys392
4,04E-04
14867 2
dys393
1,09E-03
13713 3
dys437
1,19E-03
10101 4
dys448
1,65E-03
6678 5
dys390
2,06E-03
15061 6
medium
dys385
2,30E-03
25620 7
dys19
2,32E-03
15539 8
ygatah4
2,47E-03
7709 9
dys391
2,54E-03
14935 10
dys389i
2,68E-03
13788 11
dys635
3,72E-03
7525 12
dys389ii
3,78E-03
13759 13
dys456
4,19E-03 14
dys481
4,97E-03
1744 15
dys533
5.01E-03
1730 16
dys439
5,35E-03
10096 17
dys460
6,22E-03
1717 18
dys458
6,74E-03
6677 19
dys518
1,84E-02
1556 20
fast
dyf387S1ab
1,59E-02
1804 21
dys576
1,43E-02
1727 22
dys570
1,24E-02
1426 23
dys627
1,23E-02
1766 24
dys449
1,22E-02
1617 25
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

7. Continental proportions of distinct haplotypes determined with slow, middle and fast mutating marker sets
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

8. Increased discrimination
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

9. Local Y-STR diversity differs between continents
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

10. Growth of the YHRD (2000-2014) All haplotypes Haplotypes 17 loci
Haplotypes with Y-SNPs
Haplotypes 23 loci
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

11. Count Estimates Augmented Choose the most informative haplotype format
17 loci
0/19,593
0/71,246
1.4 x 10-5
5.1 x 10-5
260/125,700
2.1 x 10-3
23 loci
9 loci
Count frequency
Choose the most informative haplotype format
in the largest available database!
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

12. Frequency estimation methods
Counting methods
Methods taking evolutionary distance between haplotypes into account
Augmented counting (1/n+1)
Counting with database inflation (Brenners κ)
Surveying method (Krawczak)
Coalescence based estimation (Caliebe)
Discrete Laplace method (Andersen)
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

13. Case-sensitive workflow (touch DNA, mixture expected)
Autosomal analysis
Mixed female/male profile
Autosomal male profile clearly identifiable
Report
National Police Database
Y chromosomal analysis
Male profile
Direct matching possible?
Database (YHRD) frequency
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

14. National Police Database
Charité Pilot study to evaluate the workflow: 40 sexual crimes with only „touch“ DNA (2011)
40 cases
8 cases
Individual male autosomale profile
10 cases
Informative YSTR Profile
22 cases
inconclusive
National Police Database
Report with
YHRD match statistics
Number of
informative profiles
doubled
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2013

15. Casework statistics since implementation of the PPY23 kit (08/2012 bis 02/2014)
160 cases
(1038 contact stains)
autosomal
No male admixture observed in the Amelogenin system
30%
Additional Y peak in Amelogenin
70%
37.5% informative for the male component
Y-chromosomal
No Y profile (11%)
Y profile observed (89%)
informative Y profile 58%
2012 – 21 Vergleichsproben
2013 – 31 Vergleichsproben
29% (=14 delicts) with Y-Profil
Single source
Y profile
(>20 Y-STRs)
39%
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

16. Case example 1: touch stain on fabric in a sexual assault case
Autosomale Analyse
Y-chromosomale Analyse
Kontaktspur, Sexualdelikt
Nur ♀-Komponente, ohne Hinweis auf ♂-Beimischung im Amelogenin
Vollständiges Y-chromosomales Einzelprofil in 23 loci
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

17. Case example 2: touch stain on fabric in a child abuse case
Yfiler + SNaPshot
9/16 Y-STRs detectable
Haplogroup I (M170)
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

18. Case example 2 (continuing): Prediction of ancestry
Haplogroup I (I2a) can also be
reliably predicted from YHRD data
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

19. Case example 3: Serial rape - linking 3 crime scenes
DYS
389I
448
389II 19
391
438
437
635
390
439
392
393
458
385ab
456
GATA H4
Case1 13 29 14 10 12 15 24 22 17
11,14 16
Case2 nd
Case 3
13+
19+
29+
14+
10+
12+
15+
22+
11,14+
16+
Consensus profile
DYS
389I
448
389II
391
437
393
458
385ab
456 13 19 29 10 15 17
11,14 16
Suspect 12 27 11 14
Exclusion of the suspect
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

20. Is NGS the future for complete resolution of male relatives ?
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

21. Next generation sequencing
10 Mb (of 24 MB NRY) sequenced
67 STRs identical
4 SNPs different
Potentially long sequence reads more variable than STRs
Xue Y, Tyler-Smith C 2009
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

22. Summary
Wider acceptance of forensic Y chromosome analysis due to a new generation of diagnostic kits with high discriminative power and much improved performance
Complete resolution of distant relatives in most populations
a proportion of close relatives can be differentiated
Adaptation of the workflow, otherwise you‘re losing evidence!
Necessity to employ statistics (LR) to evaluate the match probability
Haplotype databases accomodate the profiles of the new generation kits (
Y-STRs are included in different National DNA Databases (USA, Netherlands, Austria)
For Forensic or Paternity Use Only.
Yfiler is a trademark of Thermo Fisher Scientific and its subsidiaries.
©Charité – Universitätsmedizin Berlin, Dept. Forensic Genetics 2014

23. Disclaimers For Forensic or Paternity Use Only.
Yfiler is a trademark of Thermo Fisher Scientific and its subsidiaries.