You are on page 1of 2

Forensic Science International: Genetics Supplement Series 3 (2011) e552–e553

Contents lists available at ScienceDirect

Forensic Science International: Genetics Supplement Series
journal homepage: www.elsevier.com/locate/FSIGSS

87 DNA markers for a paternity testing: Are they sufficient?
Ilaria Carboni a, Sara Iozzi a, Anna Lucia Nutini a, Pasquale Giuseppe Macrı̀ b,
Francesca Torricelli a, Ugo Ricci a,*
a
Diagnostic Genetics Unit, Azienda Ospedaliero-Universitaria Careggi, Florence, Italy
b
Legal Medicine Institute, ASL8 Arezzo, Italy

A R T I C L E I N F O A B S T R A C T

Article history: We report a judicial paternity testing with two exclusions at D2S1338 and vWA loci. Since these results
Received 3 October 2011 suggested that the true father should be a close male relative of the tested man, mother was included into
Accepted 12 October 2011 analysis. Subjects were also typed for 24 validated STRs, 11 STRs for linkage analysis, 8 X-STRs and 30
DIPs, for a total of 87 markers. No further exclusions were found. Paternity index, taking into account
Keywords: mutation rates for D2S1338 and vWA, was 1.45  1013 (W = 0.99999999999993). The final odds that the
Paternity testing true father should be the untyped brother of the alleged father, that refused DNA profiling, was 1:192.
Kinship
ß 2011 Elsevier Ireland Ltd. All rights reserved.
STR
Relative
Forensic genetics

1. Introduction using alternative kits. Two legal sons of the alleged father (S1 and
S2) showed full compatibility at vWA and D2S1338 loci,
Paternity and kinship testing is now routinely performed by confirming father’s genotype. In order to clarify the pedigree,
using a panel of 15 STRs to obtain exclusion or strong probability of we also asked for AD mother’s (M) exhumation and we widened
paternity. However, in a small proportion of cases, this number can markers panel with most of available commercial kits, 8 X-STRs, 11
result insufficient to solve the case. This event usually happens STRs for linkage analysis mapping on chromosomes 15 and 16 and
when the mother is not available for the test or when the biological with a panel of 30 commercial DIPs, for a total of 87 DNA markers.
father is a close relative of the legal one. In these cases the test can The analysis was complicated by using samples coming from
become a tangled scenario. A large proportion of ambiguous results difficult tissues as paraffin-embedded blocks or muscle and bone,
arise in fact from unknowingly testing a first-degree relative of the that are usually degraded or contain inhibitors, reasons why
true father, usually a brother, so the exclusion rate is markedly markers with higher molecular weight did not amplify.
reduced and a paternity index using a likelihood ratio against a
random man does not apply. Less frequently, ambiguous STR
results occur from observing exclusions that may originate from 2. Materials and methods
germ-line step mutation [1]. This event occur relatively frequently
in STR loci commonly used in commercial kits; one mutation is in AD, S1 and S2 DNA samples were extracted from buccal swabs
fact expected in 2–3% of all meioses [2]. The main recourse for using BioRobotEZ1 Advanced in association with EZ11Investigator
laboratories finding such results is addiction of extra STRs to Kit. DNA from paraffin-embedded blocks, muscle and bone samples
improve the probability or, alternatively, to give unambiguous was extracted with QiaAmpDNA Micro kit and IQ-SystemTM; DNA
exclusion. was quantified with Quantifiler1. DNA samples were profiled with
Here we present a case of a woman (AD) who asserted to be a AmpFlSTR1Identifiler1Plus, AmpFlSTR1NGMTM, AmpFlSTR1Mini-
natural daughter of a deceased man (AF). The analysis of 15 FilerTM, PowerPlex116, PowerPlex116 PentaD and PentaE, Power-
commercial STRs showed two inconsistencies at vWA and Plex1ESI 17, DIPplex and ArgusX-8. We also performed analyses for
D2S1338 loci. The mismatches were reproduced and confirmed 26plex autosomal STR assay [3], eleven STR dinucleotides localized
on chromosomes 15 and 16 used for diagnosis of uniparental disomy
and for a pool of STRs used in forensic community in the past years as
* Corresponding author at: SOD Diagnostica Genetica, Azienda Ospedaliero-
F13A01, F13B, FES FPS, CD4 and LPL. CE analysis was performed on
Universitaria Careggi, Florence, Italy. Tel.: +39 055 7946204; fax: +39 055 7946200. ABI PRISM 310, 3100 and 3500 Genetic Analyzer with GeneMapper
E-mail address: ricciugoG@gmail.com (U. Ricci). v.3.2 and v.ID-X.

1875-1768/$ – see front matter ß 2011 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.fsigss.2011.10.018

who showed compatibility The judge decided for alleged father paternity.J. Am. Int. et al. X-chromosome analysis between daughter and alleged father fully matches for 8 STRs tested. In this structure and length of the tandem repeat. with paternity alternative kits. 1 (2008) 492–493.2 29–30 AF 1–2 1–2 1–3 10 12 16 15 40. / Forensic Science International: Genetics Supplement Series 3 (2011) e552–e553 e553 Table 1 Genotypes of the subjects involved in the test.3 12–14 12–12 20–25 15–15. PI calculation gave a value of 1. we considered that value was calculated including all but X-STRs and dinucleotides some mutational event should be happened in AF tumoral DNA.nist. [3] K. The final odds that the true Anyway. In a routine paternity analysis performed with 15 commercial indicating that they are likely to share the same maternal line. For conservative reasons.2–31. tissues and genotyping AF’s legal sons. Forensic excluded. any doubt was dispelled by analyzing DNA from healthy father should be the untyped uncle was 1:192. et al. Carboni et al. J. Since two Conflict of interest exclusions in a paternity testing were already described [4].2–29.2 9–13 12–13 10–12 8–10 5–10 12–13 12–15 11–11 14–14 12–12 12–12 AF 17–22 6–7 8–9 24. Brinkman. No further inconsistencies were detected. In bold the exclusions between alleged father and daughter.D. legal son 1 (S1).2 29 Daughter (AD).. mapping on chromosomes 15 and 16. 62 (1998) 1408– case. legal son 2 (S2). Genet. Hum. Suppl. The introduction of mother’s genotype just confirmed the two exclusions. Results and discussion to provide a its own DNA sample.2 10–11 10–11 14–15 6–9.2 17–19 8–8 12–17 11–11 21–22 M 14–15 30–31 10–11 11–12 14–18 9–9. STR.3 10–14 11–12 20–24 12–15. . Forensic Sci.2–29. The hypothesis that mismatches were true mutations is to take References into account: step mutations are characterized by one or two [1] B. extended the analysis for a total of 87 DNA markers. The deceased alleged father had a brother.cstl. mother (M).3 18–18 29.2 18–19 8–11 12–17 11–12 18–22 D12S391 F13A01 F13B Se33 Penta D Penta E FES FPS D4S CD4 LPL D1S D20S D6S474 D12 D9S 2364 1656 1082 GATA63 1122 AD 17–22 5–7 8–8 24.99999999999993. Balloch. allele. alleged father (AF). 3. Mutation rates in human microsatellites: influence of the repeat additions or diminutions creating an incompatibility.3 18 29.. (19 to 20 at D2S1338) and one diminution (20 to 19 at vWA) of one [2] www.2 10–11 10–11 14–15 9. the involvement of a close male relative cannot be [4] C.gov/div831/strbase. et al.3 11–11 14–14 12–17 11–12 M 16–17 5–6 8–10 26. this lymph nodes affected by Hodgkin’s lymphoma.2 10–11 10–11 14–16 6–9. with father at vWA and D2S1338 loci (Table 1). we considered the results insufficient to solve the question and we None. I.3 10–11 10–11 19–19 12–14 16–20 8–9 12–17 9–11 21–22 AF 13–16 28–31. A 26plex autosomal STR assay to aid human identity testing. Otherwise.3 11–11 10–12 17–19 12–14 15–16 9–10 11–12 9–12 21–22 S1 13–15 30–31.2 9–13 12–13 10–10 10–10 8–10 11–12 12–12 11–11 14–17 12–12 12–13 D2S1776 D10S1435 D1S1627 D3S4529 D17S1301 D1GATA113 D2S441 D77 D45 D131 D70 D6 D111 D58 D56 AD 11–13 11–14 14–14 13–15 9–12 7–12 10–11 / /+ +/+ / /+ /+ /+ /+ AF 11–12 12–14 14–14 13–16 9–11 12–12 11–11 / /+ +/+ / +/+ +/+ /+ +/+ M 13–13 11–12 13–14 13–15 12–13 7–12 10–14 / +/+ /+ / / /+ /+ /+ D118 D92 D93 D99 D88 D101 D67 D83 D114 D48 D124 D122 D125 D64 D81 AD +/+ / /+ /+ / +/+ +/+ / /+ +/+ /+ +/+ /+ +/+ /+ AF +/+ / /+ / /+ /+ /+ /+ / /+ /+ /+ +/+ /+ /+ M +/+ / /+ +/+ /+ /+ /+ / /+ /+ /+ /+ /+ /+ / D136 D133 D97 D40 D128 D39 D84 D15S1007 D15S120 D15S128 D15S516 D15S1012 D15S978 D16S3103 D16S423 AD /+ /+ /+ / /+ / /+ 1–3 2–3 1–2 1–1 1–1 1–1 2–2 1–1 AF /+ /+ /+ / / / / 2–3 1–2 1–3 1–2 1–2 1–1 2–3 1–1 M +/+ /+ / / /+ / +/+ D16S415 D16S3046 D15S205 DXS8378 HPRTB DXS7423 DXS7132 DXS10134 DXS10074 DXS10101 DXS10135 AD 1–1 2–2 2–3 10–11 12–14 15–16 15–15 34–40. Ser.3 10–11 12–12 20–25 12–15 15–17 8–11 12–13 11–12 18–21 S2 13–13 30–31. two inconsistencies at vWA and D2S1338 loci between a Thus. 54 (5) (2009) 1008–1015. the two inconsistencies are just characterized by one addition 1415. Since AF typing came from paraffin-embedded probability W = 0. who refused Sci. Reporting paternity testing results when 2 exclusions are encountered..: Genet. incorporating vWA and D2S1338 mutation rates into daughter and alleged father were detected and confirmed with Familias.2 9–12 12–13 11–12 8–9 5–10 11–13 15–17. D8S1179 D21S11 D7S820 CSF1P0 D3S1358 TH01 D13S D16S D2S1338 D19S433 vWA TPOX D18S51 D5S818 FGA 317 539 AD 13–15 31–31.2–26. J.45  1013. Hill.3–9.2 10–11 10–10 15–16 6–9.