RESUMO
In a case of the death of a 7-year-old boy, the police investigations revealed a possible dog attack contrary to the witness testimonies. DNA investigations were carried out from hairs, saliva and bloodstains with 10 canine-specific STR loci by the use of fluorescently labelled multiplex PCR and the ABI PRISM 310 genetic analyzer. The analysis of one hair sample revealed one allele deviation from the profile of the putative Rottweiler perpetrator possibly caused by a mutation. The PCR fragments in question at the PEZ20 locus were sequenced and compared with the alleles detected in the Hungarian canine population and identified on a repeat number basis. The allele frequencies were determined based on typing of 242 genetically independent canine individuals from 72 breeds. The results suggested that two of the canine individuals could be the perpetrators.
Assuntos
Cães/genética , Medicina Legal , Cabelo/química , Sequências de Repetição em Tandem , Animais , Mordeduras e Picadas , Manchas de Sangue , Criança , Mapeamento Cromossômico , Frequência do Gene , Humanos , Masculino , Mutação , Reação em Cadeia da Polimerase , Saliva/química , Especificidade da EspécieRESUMO
Several animal carcasses were found in the paddocks of a Hungarian County Zoo during 1 week. The 14 animals killed were thought to be the victims of a dogfight training. The primary suspect was the security guard of the Zoo with his guard dogs. DNA tests were carried out on hairs and bloodstains and 10 canine-specific STR loci were analysed by fluorescently labelled multiplex PCR using the ABI PRISM 310 Genetic Analyzer. The results confirmed that the killer was a single animal and all of the guard dogs were excluded.
Assuntos
Medicina Legal , Repetições de Microssatélites/genética , Polimorfismo Genético , Bem-Estar do Animal , Animais , Crime , Cães , Marcadores Genéticos , Cabelo/citologia , Hungria , Patologia Veterinária , Reação em Cadeia da PolimeraseRESUMO
A multiplex reaction for the eight STR loci D3S1358, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820 was used to generate allele frequency databases for two Hungarian population samples, Caucasians from the Budapest area and Romanies from Baranya county. During the analysis two intermediate-sized alleles and a sequence variant allele were observed at the D7S820 locus. All three types of allelic variants were found to have modifications in the same block of a (T)9 stretch located within the 3' flanking region of each allele, which may indicate a possible higher mutation rate of this (T)9 block. For the loci D3S 1358 and D7S820 the Romany population database showed departures from Hardy-Weinberg equilibrium. The forensic efficiency values for the Romany population were slightly different from those found in the Hungarian Caucasian population. Comparing the allele frequency values by G-statistic, calculating the F(ST) indices and with the pair-wise comparisons of interpopulation variance, the two Hungarian populations could be distinguished using data from the eight STR loci.
Assuntos
Medicina Legal , Genética Populacional , Roma (Grupo Étnico)/genética , Sequências de Repetição em Tandem/genética , População Branca/genética , Albânia , Interpretação Estatística de Dados , Eletroforese , Feminino , Frequência do Gene , Variação Genética , Genótipo , Humanos , Hungria , Masculino , Mutação , Reação em Cadeia da Polimerase , Polimorfismo Genético , Software , IugosláviaRESUMO
A collection of eight STR loci (D3S1358, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820) was used to generate allele frequency databases for two Hungarian population samples, Caucasians from the Budapest area and Romanies from Baranya county. During the analysis two intermediate sized alleles and a sequence variant allele were observed at the D7S820 locus. All three types of allelic variants were found to have modification (deletion, insertion, transversion) in the same block of a (T)(9) stretch located within the 3' flanking region of each allele, which may indicate a possible higher mutation rate of this (T)(9) block. For the loci D3S1358 and D7S820 the Romany population database showed departures from Hardy-Weinberg equilibrium. The forensic efficiency values for the Romany population were slightly different from those found in the Hungarian Caucasian population. Comparing the allele frequency values by G-statistic, calculating the F(st) indices and with the pairwise comparisons of inter-population variance, the two Hungarian populations could be distinguished using data of the eight STR loci.
Assuntos
Impressões Digitais de DNA/métodos , Frequência do Gene/genética , Repetições Minissatélites/genética , Roma (Grupo Étnico)/genética , População Branca/genética , Consanguinidade , Bases de Dados Factuais , Variação Genética/genética , Humanos , Hungria , Paternidade , Polimorfismo Genético/genética , Estudos de AmostragemRESUMO
A set of seven Y-chromosomal STR loci (DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393) with the addition of the bilocal marker DYS385 was used to generate male-specific haplotype databases for two Hungarian population samples, Caucasians from the Budapest area and Romanies from Baranya county. At the locus DYS385 three types of intermediate sized alleles were detected in six males. The presence of a (GA) dinucleotide, probably due to an (AA) deletion in the second (GAAA) repeat of the polymorphic repeat region leads to an intermediate allelle 17.2. The intermediate alleles 17.-1 and 18.-1 with the consensus repeat structure of (GAAA)17 and (GAAA)18, respectively, were found to lack a T in the same (T)7 stretch located within the 3' flanking region of each allele. The forensic efficiency values for the Romany population were significantly lower than those found in the Central Hungarian and other non-isolated Causasian populations, which may imply a possible common paternal ancestry of some haplotypes in the Romany sample. With pairwise comparisons of inter-population molecular variance, the two populations analyzed here and an Italian population sample, could be clearly distinguished using the seven monoclonal Y-STRs. A sizing precision of < or = 0.14 nucleotide standard deviation was obtained with capillary electrophoresis carried out on an ABI Prism 310 Genetic Analyzer. Objective and accurate genotyping is thus possible using an internal size standard with a high density of fragments.
Assuntos
Alelos , Impressões Digitais de DNA/métodos , Repetições de Dinucleotídeos/genética , Marcadores Genéticos/genética , Haplótipos/genética , Repetições Minissatélites/genética , Polimorfismo Genético/genética , Roma (Grupo Étnico)/genética , População Branca/genética , Cromossomo Y/genética , Análise de Variância , Bases de Dados Factuais , Eletroforese Capilar , Variação Genética/genética , Genótipo , Humanos , Hungria , Itália , Masculino , Reprodutibilidade dos TestesRESUMO
Population data were generated for the STR systems HUMFIBRA and HUMD21S11 for a Hungarian Caucasian population sample residing in Baranya County, Hungary (127 unrelated individuals). The loci were coamplified using a fluorescence based PCR method and were typed automatically. For both loci 12 different alleles could be found including some variants. No deviations from Hardy-Weinberg expectations were observed. Both loci proved to be highly discriminating and valuable polymorphisms for forensic analyses.
Assuntos
Alelos , Marcadores Genéticos , Genética Populacional , Sequências Repetitivas de Ácido Nucleico , População Branca/genética , Medicina Legal , Humanos , Hungria , Reação em Cadeia da Polimerase/métodos , Polimorfismo GenéticoRESUMO
A population study of Hungarian Romanies was carried out on the STR loci HumLPL, HumF13B, HumFES and HumF1301. There was little evidence for association of alleles within/between the four STR systems. Allele frequency distributions were significantly different between the Romany and the previously reported Central Hungarian population databases. Population differentiation was estimated by computing F- and phi-statistics as well as frequency estimate differences of individual phenotypes for these two population samples. The results suggest that the population structure may have an effect on the interpretation of forensic DNA evidence in Hungary. Phylogenetic tree reconstruction with six populations from three major ethnic groups revealed a relatively distant genetic relationship of the Baranya Romanies with other Caucasian populations.
Assuntos
Alelos , Mapeamento Cromossômico , Genética Populacional , Roma (Grupo Étnico)/genética , Sequências de Repetição em Tandem , Frequência do Gene/genética , Humanos , Hungria , Análise de Sequência de DNARESUMO
A panel of monoclonal antibodies was generated against the urea-based hapten N-(2-N-chloroacetylaminobenzyl)-N'-4-chlorophenylurea as a tool for building up sensitive immune assays to detect urea derivatives and to screen them for catalytic antibodies (Abs). Eleven hybridomas were obtained that produced Abs reactive to the hapten. All Abs were of IgG class. Cross reactivities of the Abs to different haptens were examined, especially to a possible transition-state analog. Only four of the hybridomas (R2-DA10/F7, R2-GE7/H2, R2-HC2/A5, R2-HD6/F7) produced Abs crossreactive with the transition-state analog. From the 11 hybridomas, hybridoma B76-BF5 was chosen for further characterization. Compared to the other Abs, B76-BF5 showed the strongest binding and had a rather restricted specificity. These Abs could be used to build up a sensitive enzyme immunoassay for the detection of the hapten. All Abs were screened for crossreactivity with the pesticides monuron and diuron. No reactivity could be detected. In addition, the nucleotide sequences of the variable light and heavy chain genes of the similarly reactive Abs B76-BF5, B76-BB3, R2-DA10/F7, and R2-GA6/G3 were determined to clarify whether structure and binding specificity of these Abs showed any correlation.
Assuntos
Acetamidas/imunologia , Anticorpos Monoclonais/isolamento & purificação , Praguicidas/imunologia , Compostos de Fenilureia/imunologia , Ureia/análogos & derivados , Ureia/imunologia , Sequência de Aminoácidos , Animais , Reações Cruzadas , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência MolecularRESUMO
Allele frequencies of the three STR systems HUMLPL, HUMF13B, and HUMF13A01 were determined in a Hungarian population sample of 223 unrelated Caucasian individuals. All loci met Hardy-Weinberg expectations and there was no evidence for association of alleles among the three STR loci. In addition, little evidence was found for departures from expectations of independence between any of the three STR and other previously investigated microsatellite polymorphisms.
Assuntos
Genética Populacional , Repetições de Microssatélites/genética , Alelos , Frequência do Gene/genética , Homozigoto , Humanos , HungriaRESUMO
A population study of Hungarian Romanies was carried out for the STR loci HumVWFA31, HumTH01, HumTPOX, and HumCSF1PO. After multiplex PCR amplification semi-automatic DNA profiling was performed using an ALF DNA sequencer. At the loci investigated there was little and no evidence for departures from Hardy-Weinberg expectations and linkage equilibrium, respectively. The allele sizing accuracy of the ALF DNA sequencer was increased to a high level (99.97% on average) by applying external and internal markers. Allele frequency distributions of the STR loci, with one exception, were significantly different between the Romany and other Hungarian population databases. On the other hand, however, only small differences in frequencies of individual phenotypes were found.
Assuntos
DNA/genética , Marcadores Genéticos/genética , Genética Populacional , Reação em Cadeia da Polimerase , Roma (Grupo Étnico)/genética , Adulto , Alelos , Feminino , Frequência do Gene/genética , Homozigoto , Humanos , Hungria , MasculinoRESUMO
The authors survey the application of polymerase chain reaction (PCR)-based DNA polymorphisms in the Hungarian forensic practice. The combined application of the presented 17 PCR-based sequence- or length-polymorphic DNA systems to criminal cases gives the power of individualization to the hand of the forensic scientist. The joint application of these genetic markers to disputed paternity cases enables the verification of paternity for an unexcluded man with the highest legal category, namely "paternity practically proved". The investigation of sex-chromosome linked STRs and/or the analysis of mitochondrial DNA (mtDNA) D-loop region is useful in solving most of the problematic cases. The highlighted advantages of PCR over restriction fragment length polymorphism (RFLP)-based forensic DNA analyses clearly explain the overwhelming spread of PCR-based methods in the Hungarian forensic practice.
Assuntos
DNA/análise , Medicina Legal , Paternidade , Reação em Cadeia da Polimerase , Humanos , Hungria , Masculino , Polimorfismo GenéticoRESUMO
Data have shown that on the internal surface of reused haemodialysis membranes complement elements of plasma proteins, cells and cellular debris from the patient's blood are deposited. The aim of our study was to identify the effect of reuse on the haemodialysis membrane surface. Original and reused haemodialysis membranes were studied by the methods of light, transmission and scanning electron microscopy, immuno-histochemistry and X-ray spectrum element analysis. With reuse the membrane surface becomes uneven, and its continuity may break. Plasma protein and cellular elements are deposited on the damaged surface following continued reuse. Presence of IgM deposists were confirmed by immunohistochemical observation. Moreover, following reuse the X-ray spectrum analysis verified structural changes in the haemodialysis tube compared to the control material; accumulated aluminium, silicon, phosphorus, sulphur and chlorine were detected on the wall of the dialysis membrane.
Assuntos
Celulose/análogos & derivados , Membranas Artificiais , Diálise Renal/normas , Alumínio/metabolismo , Celulose/química , Celulose/metabolismo , Cloro/metabolismo , Eletroforese em Gel de Poliacrilamida , Reutilização de Equipamento , Humanos , Imuno-Histoquímica , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Fósforo/metabolismo , Silício/metabolismo , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Enxofre/metabolismo , Propriedades de SuperfícieRESUMO
Hungarian population data for the loci LDLR, GYPA, HBGG, D7S8, Gc, HLA-DQA1, and D1S80 were generated. The genotype frequency distributions for the loci do not deviate from Hardy Weinberg expectations. Furthermore, there was little evidence for departures from expectations of independence between the loci. Using a test for homogeneity all the loci were similar between two Hungarian population samples and only the HLA-DQA1 locus was statistically different between Hungarians and US Caucasians. There generally would be little forensic differences, whether a Hungarian or a US Caucasian database was used, for estimating multiple locus profile frequencies for the seven PCR-based loci.
Assuntos
DNA/análise , Genética Populacional , Reação em Cadeia da Polimerase , População Branca/genética , Frequência do Gene , Genótipo , Humanos , Hungria , Funções VerossimilhançaRESUMO
Population data studies for HLA-DQ alpha and PM loci (LDLR, GYPA, HBGG, D7S8, GC) were carried out on a Hungarian Caucasian population sample of 163 unrelated individuals. Whereas the observed PM allele frequencies were similar to those reported for Caucasians, significant differences were found for HLA-DQ alpha between the Hungarian and some Caucasian population data. All six loci meet Hardy-Weinberg expectations and there is no evidence for association between any pairs of loci.
Assuntos
Mapeamento Cromossômico , Marcadores Genéticos/genética , Genótipo , Reação em Cadeia da Polimerase , Alelos , Frequência do Gene/genética , Triagem de Portadores Genéticos , Genética Populacional , Antígenos HLA-DQ/genética , Cadeias alfa de HLA-DQ , Homozigoto , Humanos , Hungria , Polimorfismo Genético/genéticaRESUMO
We present a Hungarian population study for six tetrameric short tandem repeat (STR) loci employing multiplex PCR amplification, electrophoresis of the PCR products in DNA sequencing gels and subsequent detection of allelic fragments by silver staining. The loci were HUMVWFA31, HUMTH01, HUMCSF1PO, HUMFES/ FPS, HUMTPOX, and HUMHPRTB. All loci met Hardy-Weinberg expectations in the examined Hungarian Caucasian population sample (N = 223 individuals). In addition, there was no evidence for association of alleles among the five autosomal loci HUMVWFA31, HUMTH01, HUMCSF1PO, HUMFES/FPS, and HUMTPOX.
Assuntos
Alelos , Impressões Digitais de DNA , Etnicidade/genética , Sequências Repetitivas de Ácido Nucleico/genética , Feminino , Frequência do Gene , Humanos , Hungria , Masculino , Reação em Cadeia da Polimerase/métodos , População Branca/genéticaRESUMO
Population data studies for TH01 and VWA were carried out on a Hungarian caucasian population sample of 244 unrelated individuals. We found 7 and 8 different alleles for TH01 and VWA, respectively. Using a high resolution/non-denaturing horizontal PAGE system no variations in the mobility of alleles were detected except for the allele 14 in VWA. The allele frequencies observed are similar to those reported in the literature for European caucasians. The genotype distributions meet Hardy-Weinberg expectations for both STR systems.
Assuntos
Alelos , Frequência do Gene , Sequências Repetitivas de Ácido Nucleico/genética , Distribuição de Qui-Quadrado , Impressões Digitais de DNA , Marcadores Genéticos , Genótipo , Humanos , Hungria , Modelos Genéticos , Reação em Cadeia da Polimerase , Reprodutibilidade dos TestesRESUMO
Population data studies for D1S80 and ApoB were carried out on a caucasian population sample from Hungary of 229 and 222 unrelated individuals, respectively. We observed 26 different alleles for D1S80 and 13 for ApoB. The allele frequencies found are similar to those reported in the literature for European caucasians. No evidence of significant deviations from Hardy-Weinberg equilibrium were observed for both AMPFLP systems.