0000000000335359
AUTHOR
U. Schacker
Application of DNA Polymorphisms in Paternity Testing in Germany: Solution of an Incest Case Using Bacteriophage M13 Hybridization with Hypervariable Minisatellite DNA
More than 25 blood, serum, and enzyme polymorphisms have been introduced into paternity testing in Germany in recent years (Rittner, 1975). If a “no” decision is defined by exclusion, and a “yes” decision requires a probability of 99.73 % or more, more than 90 % of court cases can be solved in this respect. A few cases not being clarified by a standard expertise include: 1) Cases with more than one alleged man if the men and/or the mother and the men are related. 2) Some cases where the putative father is deceased, and neither the parents nor the legitimate offspring are available for the study. 3) Cases where possible exclusion in a given polymorphic system interferes with an overall evide…
Reliability of Restriction Enzyme Digestions of Genomic DNA for the Generation of DNA Fingerprints
Since minisatellite DNA probes are used for the detection of hypervariable loci in eucaryotic genomes [1] the application of so called DNA fingerprints and DNA technology itself in paternity testing and forensic casework is critically discussed ([3]; Brinkmann et al., this volume). A particular problem is the possibility of obtaining partially digested genomic DNA in casework after treatment with restriction enzymes leading to inconclusive or even false results. This is even more important when multilocus DNA probes are used, since the total number of fragments in a given person is not known in advance. But also with single locus probes, where only two allelic fragments are usually detected…
Minisatellite DNA Probe MZ 1.3: Application in Paternity Testing and Estimate of the Number of Genetic Loci
The use of hypervariable DNA minisatellite probes recognizing repetitive genomic DNA sequences has become a valuable and powerful tool in paternity testing as well as in forensic stain analysis (Jeffreys et al. 1985, 1987; Werrett et al. 1988). It has been shown that bacteriophage Ml3 DNA can also be used to obtain hypervariable DNA restriction fragment patterns in humans and other species (Vassart et al. 1987). To obtain more informative and specific fragment patterns for the DNA ‘fingerprint’ analysis in man, we have used Ml3 DNA as a probe to screen a human genomic library. Thus, we have isolated the minisatellite DNA probe MZ 1.3 (Schacker et al., in press). MZ 1.3 is a 1.9 kb fragment …
Quantitative and qualitative analysis of DNA extracted from postmortem muscle tissues
DNA extracted from 33 postmortem muscle specimens was analyzed using MZ 1.3, a hypervariable minisatellite probe, as well as locus-specific minisatellite probes (g3, MS1 and MS43). After storage at -25 degrees C for 10 months, DNA from all the samples was partially (approximately 21% of total DNA) degraded even when autopsy was performed 1 day postmortem. However, more than 90% of DNA samples up to at least 3 days postmortem were suitable to obtain good restriction fragment length polymorphism (RFLP) patterns. When small strips of specimen were stored for 8 days at room temperature in moist chambers, approximately 42% of total DNA was degraded. Only 30% of these DNA samples still showed goo…
Old meets new: Comparative examination of conventional and innovative RNA-based methods for body fluid identification of laundered seminal fluid stains after modular extraction of DNA and RNA.
Abstract The knowledge about the type of the body fluid/tissue that contributed to a trace can provide contextual insight into crime scene reconstruction and connect a suspect or a victim to a crime scene. Especially in sexual assault cases, it is important to verify the presence of spermatozoa. Victims often tend to clean their underwear/bedding after a sexual assault. If they later decide to report the crime to the police, in our experience, investigators usually do not send laundered items for DNA examination, since they believe that analysis after washing is no longer promising. As not only the individualization of traces on laundered items could be important in court, but also the type…
Paternity Analysis Using the Multilocus DNA Probe MZ 1.3
The multilocus minisatellite DNA probe MZ 1.3 detects hypervariable restriction fragment patterns in genomic DNA of man and animals. It can be used for segregation analysis in cases of disputed paternity (Schacker et al., 1991; Rittner et al., 1991a), for identification purposes in forensic medicine and stain analysis (Ogata et al., 1990; Rittner et al., 1991b), as well as in animal breeding for pedigree analysis and verification of inbred strains (Hins & Gruber, 1991). Hypervariable fragment patterns can be generated by using frequently cutting restriction enzymes, e.g. Hinf I, Hae Ill, Msp I, Mbo I, and Rsa I. A non-radioactive system using the digoxigenin antidigoxigenin system may be us…
Non-isotopic DNA fingerprint analyses with the minisatellite probe MZ1.3
RFLP analyses with minisatellite probes yield highly informative individual specific banding patterns (genetic fingerprints) (Jeffreys et al.). Because of the complexity of the pattern and marked differences in band intensities a good band resolution and high sensitivity of the probe are essential. By reason of the latter requirement to date most of the RFLP-studies are performed with radioactive probes. Although non-isotopic labels have been introduced into fingerprint analyses (Schafer et al.; Medeiros et al.) 32P-labeled probes are still superior with respect to sensitivity. Our approach to increase the specific signal intensity makes use of an amplification of the number of probe molecu…
Possible association of sudden infant death with partial complement C4 deficiency revealed by post-mortem DNA typing of HLA class II and III genes
Based on evidence of an increased rate of respiratory infections in sudden infant death (SID) infants as well as the observation of familial occurrence, we analysed in a retrospective study class II and class II genes of the major histocompatibility complex in 40 cases of SID by Southern blot analysis of DNA obtained post mortem from tissue samples. In 24 cases, the parents were interviewed and confirmatory human lymphocyte antigen (HLA) and DNA typing was carried out. Using HLA-DR beta and -DQ beta probes, no evidence of an abnormal HLA-DR frequency distribution in SID infants was detected (P = 0.97). Using DNA probes for the tandemly arranged complement C4 and steroid 21-hydroxylase genes…
Isolation of the DNA minisatellite probe MZ 1.3 and its application to DNA ‘fingerprinting’ analysis
Abstract A minisatellite probe, MZ 1.3, detecting hypervariable fragment patterns was isolated from a human genomic library. A repetitive sequence of 27 bp length was identified which is contained in the probe approx. 40 times. The MZ 1.3 repeat shows variable homology of 53–73% to the repetitive sequence of the protein III gene of the bacteriophage M13 genome. Polymorphic restriction fragment patterns were found with MZ 1.3 using the enzymes Hinf I, BstN I, Hae III, Mbo I, PstI PvuII , and Rsa I. An average of 18 polymorphic fragments was observed using Hinf I as enzyme. The band sharing frequency after Hinf I digestion among unrelated individuals was determined to be 23.8 ± 7.2%. An examp…
DNA Fingerprinting as a Tool of Paternity Testing in Germany
Soon after the discovery of the ABO blood groups by Landsteiner their forensic application was suggested by Max Richter at the 74. Versammlung Deutscher Naturforscher und Arzte at Karlsbad (Karlovy Vary) in 1902 [1], Landsteiner’s Nobel lecture “Uber individuelle Unterschiede des menschlichen Blutes” in 1930 [2] provided a landmark on the path towards the discovery of so-called hypervariable minisatellite regions of human DNA by Alec Jeffreys in 1985 [3]. Since the individual patterns of any human being seemed to be unique it is well understood that this achievement was immediately welcomed with great enthusiasm by the scientific community and the public opinion. Soon thereafter, however, s…