ABSTRACT
<p><b>OBJECTIVE</b>To establish a nine X-chromosome short tandem repeats (X-STR) loci multiplex PCR method and study the polymorphism of the 9 X-STR loci,and to determine its application in kinship tests for forensic medicine.</p><p><b>METHODS</b>A fluorescent multiplex PCR that simultaneously amplifies 9 X-STR loci, i.e. DXS7133, DXS981, DXS7424, DXS6789, DXS7132, GATA165B12, DXS101, GATA31E08 and DXS10011, were set up. PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer with GeneMapper ID 3.1 analysis software.</p><p><b>RESULTS</b>When 251 unrelated male and 112 unrelated female individuals from southern China were tested, 111 alleles were detected. The power of discrimination in females was 0.5837-0.9959. Mean exclusion chance for X-STR in standard trios with daughters was 0.4072-0.9511. Polymorphism information content was 0.4481-0.9531.</p><p><b>CONCLUSION</b>The results demonstrate that the 9 loci in the multiplex system provide high polymorphism information, and the multiplex system provides a fast technology for forensic identification and paternity testing. The X-STR multiplex system can complement the analysis of AS-STR and Y-STR efficiently.</p>
Subject(s)
Female , Humans , Male , Alleles , Asian People , Genetics , China , Chromosomes, Human, X , Genetics , Microsatellite Repeats , Polymerase Chain Reaction , Methods , Polymorphism, GeneticABSTRACT
Objective Estimate the paternity exclusion probability for 15 short tandem repeats (CODIS set plus Penta D and Penta E) in duo paternity tests. Methods 644 random individuals were paired to con-struct 207046 fictitious duo families. Cases of non-exclusion or with only one exclusionarylocus were count-ed. Results 27 cases could not be excluded by the 15 STR loci. The combined probability of exclusion was 0.999869. Number of cases showing only one exclusionary locus was 384 (0.185%). Conclusion To con-firm relationship in duo cases with only 15 STRs should be careful. Extensive genetic markers is needed to discriminate one locus exclusion or mutation.
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【Objective】 To understand the polymorphism of D13S63 1 in Guangdong Han population. 【Method】 Short tandem repeat (STR) locus D13S63 1 was analyzed by means of polymerase chain reaction, following discontinuous el ectrophoresis system. 【Result】 Among 227 unrelated individuals from Guangdong Han population, 6 alleles rang from 197～217 bp could be observed. The most com mon allele with a frequency 0.2907 is allele 201 bp, the rare allele is allele 197 bp with a frequency 0.0903. The heterozygosity, the power of discrimination and the exclusion chance in paternity case were 0.7885，0.9231, 0.5543, respect ively. Segregation studies reveal that D13S631 inherit in Medel's Law. 【Conclus ion】 The result shows that locus D13S631 is highly informative and suitable fo r forensic application.
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Objective To investigate the polymorphism of DXS6854 locus in Guangdong Han population. Methods The DXS6854 locus was analyzed by PCR following polyacrylamide gel electrophoresis and silver staining. Results Among 189 females and 230 males from Guangdong Han population, 8 alleles were observed with frequencies ranging from 0.0026 to 0.4522. Exact tests demonstrated genotype frequencies in females had no departure from Hardy- Weinberg equilibrium. The discrimination powers for female and male original samples were 0.8633 and 0.7012, respectively. When both mother and her daughter were tested, the probability of excluding a random man as a father was 0.6712. Conclusion The DXS6854 locus is appropriate for individual identification and paternity testing involving a female child.
ABSTRACT
By analysing the 300 cases of paternity test in our lab in the recent year, we have demonstrated that mutation rates in STR loci are very high. The experimental steps include extracting DNA by chelex 100, amplification of DNA, polyacrylamide gel electrophoresis, silver staining, DNA typing by standard samples. The STR mutations in 11 cases among 300 paternity test cases were found, induding D11S554 locus6 cases, D19S253 locus2 cases, SE33 locuslcase, D12S391 locus 1 case, D13S631 locus 1 case. The results indicated that high mutation rates in STR loci should be considered when STR genotyping are applied to paternity test.
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To investigate haplotype distribution of the Y-chromosome STRs (DYS19, DYS390 and DYS389 locus) in Han population in Guangdong area. The STRs were typed by poplymerase chain reaction followed by discontinuous PAGE system. Among 130 unrelated males, 81 different haplotypes were observed, 52 out of them were found only one time. The haplotype genetic diversity, discrimination power and non-father exclusion chance are 0. 9989, 0. 9824, 0. 9824, respectively. The high informative haplotypes make these STRs useful for the forensic individual identification and paternity testing.
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Objective To study the validation of fluorescent multiplex microsatellite amplification technique for use in the caseworks of umbilical blood cell transplantation monitor. Methods Six post-transplant recipients of umbilical cord blood cell transplantation were monitored by analyzing microsatellite DNA loci. DNA samples were amplified using a fluorescent labeling primers multiplex amplification system of 15 microsatellites markers, followed by typing on a DNA automated sequencer. Recipients' or donors' microsatellites DNA profiles were compared before and after transplantation. Results All recipients and donors exhibited different DNA profiles. Without reference samples of pre-transplant or donors, the changes of the 15 microsetellites genotypes of the post-transplant recipients still could be analyzed. The recipient type turned to donor type was observed over time. Conclusion Under the condition of using multiplex amplification of the 15 microsatellites to monitor the umbilical blood cell transplantation, reference sample of pre-transplant or donor did not need to be detected simultaneously.
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Objective To evaluate the cause which leads to the allelic dropouts at D8S1179 locus while performing paternity testing with the AmpFlSTR Profiler Plus kit. Methods A singleplex amplification system for D8S1179 locus (GenBank Accession No. G08710) was used to verify the typing results by using the AmpFlSTR?Profiler plus kit. Dropout alleles were then sequenced. Results G to A transition was identified at the position of the 147th base of the GenBank sequence. The frequency of the G to A transition among the Chinese population was 0.50 X 10"2 (10 out of 2013 unrelated individuals). Conclusion The G to A transition may be located at the binding site of one of the primers of the AmpFlSTR?Profiler Plus kit. It is suggested that the G to A transition might be the cause of the allelic dropout at the D8S1179 locus.
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Objective To investigate the STR typing discordance between different typing methods.Methods Genotypes of 13 routine forensic STR loci in DNA samples from 100 individuals were typed by using singleplex polyacrylimide gel electrophoresis silver stain system and Power Plex16 System,respectively.The typing results between these two systems were compared.Results One genotype discordance was observed at D8S1179 locus in a DNA sample.The genotypes was 12/14 in snigleplex system and 12/15 in Power Plex16 System.Conclusion Different STR typing systems may result in different genotyps from the same DNA sample.