Genetic polymorphisms of 19 X-STR loci for forensic application in China’s three ethnicities / 基础医学与临床

Objective To investigate the genetic data of the 19 X-STR loci in three ethnicities of China ( Han, Gelao,Miao) and to evaluate the application in forensic science.Methods The DNA samples of unrelated individ-ual in Han (n=308), Gelao (n=398), Miao (n=323) ethnicities were amplified using MicroreaderTM19X ID System kit, and the PCR products were analyzed by electrophoresis through 3500XL genetic analyzer. The fragment sizes of alleles were taken subsequently by GeneMapper? ID-X. Allele frequencies and national genetics parameters of the 19 X-STR were analyzed by statistics. The allele frequencies were compared among the three nationalities and were compared with available data of other Han ethnicities from different regions. Results After the Bonferroni correction at a 95% significance level, no significant departures from the Hardy-Weinberg equilibrium was observed. Linkage disequilibrium test showed no significant allelic association between all 19 X-STR loci after Bonferroni’s correction. The cumulative discrimination power in females and in males were greater than 0.999 999 999 99 and 0.999 999 999 94,respectively. The combined power of exclusion in trios and in duos were greater than 0.999 999 999 36 and 0.999 999 52,respectively. The p values,calculated throuth Arle-quin v3.5 software,there were significantly different as detected at loci of X-STR among the different nationalities. Conclusions This panel of X-STR is highly polymorphic in China’s three ethnicities and can be served as a supple-mentary to the current STR system for individual identification.

Genetic Polymorphisms of 18 Autosomal STR loci in Changsha Han Population / 法医学杂志

OBJECTIVES@#To investigate the genetic polymorphisms of 18 autosomal short tandem repeats （STR） loci in Changsha Han population, and explore the population genetic relationships and evaluate its application value in forensic medicine.@*METHODS@#The DNA of 2 004 unrelated individuals in Changsha Han population were amplified using Goldeneye®DNA ID System BASIC, and the PCR products were analyzed by electrophoresis using 3130xl genetic analyzer. The fragment sizes of alleles were analyzed subsequently by GeneMapper® ID v3.2. The frequency data and forensic genetic parameters ［observed heterozygosity （Ho）, expected heterozygosity （He）, power of discrimination （DP） and polymorphic information content （PIC）］ of 18 STR loci were statistically analyzed. Total probability of discrimination （TDP）, probability of exclusion in trio cases （PEtrio） and probability of exclusion in duo cases （PEduo） were calculated by Cervus 3.0. Hardy-Weinberg equilibrium and linkage disequilibrium of the loci were detected by Arlequin v3.5. The results were compared with the available data of other populations from different races and regions.@*RESULTS@#The power of discrimination （DP）, and the polymorphic information content （PIC） of each locus of Changsha Han population ranged from 0.783 6 to 0.987 9 and 0.549 4 to 0.914 5, respectively. The TDP, cumulative probability of exclusion in trio cases （CPEtrio） and cumulative probability of exclusion in duo cases （CPEduo） were 0.999 999 999 999 999 999 999 865 2, 0.999 999 979 and 0.999 988 325, respectively. According to the Nei's DA genetic distance, the genetic distance between Changsha Han and Hunan Han populations was the smallest （0.014 1）, while it was the largest （0.041 8） between Changsha Han and Xinjiang Kazakh populations.@*CONCLUSIONS@#The 18 STR loci shows abundant genetic polymorphisms in Changsha Han population. The study of genetic diversity among different populations has an important meaning for the research of their origins, migrations and their relationships.

Genetic Polymorphisms of 27 Y-STR Loci in Dongxiang Population of Gansu Province / 法医学杂志

OBJECTIVES@#To investigate the genetic polymorphisms of 27 Y-STR in Dongxiang population of Gansu province, and to explore the population genetic relationship and the value of forensic application.@*METHODS@#The genotyping of 27 Y-STR loci in 526 unrelated male individuals in Dongxiang population of Gansu province were detected by STRtyper-27Y kit. The allele frequencies and haplotype diversity were also calculated. Combining with other genetics data of 14 loci in same populations, which have been published at home and abroad, the genetic distance and clustering relationship in Dongxiang population of Gansu province were calculated.@*RESULTS@#Totally 55 haplotypes were found in the DYS385a/b biallelic loci, 39 haplotypes in DYF387S1 loci, and 4-16 alleles in the rest 23 single copy STR loci. The GD value was from 0.453 9 （DYS391） to 0.957 5 （DYS385a/b）. Totally 471 haplotypes were observed in 27 Y-STR loci in 526 individuals, and the value of haplotypes diversity was 0.999 5. The genetic distance between Dongxiang and Tibetan populations of Gansu province was the closest （0.068 2）, while it was the longest between Dongxiang population in Gansu province and Han population in Henan province （0.084 7）. The result of dimensional analysis established upon the genetic distance was basically matched with that of the cluster analysis.@*CONCLUSIONS@#The 27 Y-STR loci show a high genetic polymorphism in Dongxiang population of Gansu province, which has significance for the Y-STR database establishment, population genetics study and forensic practice.

Forensic Application of 16 X-STR Loci in Henan Han Population / 法医学杂志

OBJECTIVES@#To investigate the genetic data of 16 X-STR loci in Henan Han population and to assess the application value in forensic science.@*METHODS@#The DNA of 326 unrelated individuals in Henan Han population were amplified using Goldeneye™ DNA identification system 17X kit, and the PCR products were analyzed by electrophoresis through 3130xl genetic analyzer. The fragment sizes of alleles were analyzed subsequently by GeneMapper® ID-X. Allele frequencies and population genetics parameters of 16 X-STR loci were analyzed statistically and compared with the available data of other Han populations from different regions.@*RESULTS@#Among the 16 X-STR loci, DXS6800 were found to be moderately polymorphic and the other 15 X-STR loci were highly polymorphic. The cumulative discrimination power in females and males were 0.999 999 999 999 992 and 0.999 999 996 577 712, respectively. The combined power of exclusion in trios and in duos were 0.999 999 971 and 0.999 992 574, respectively.@*CONCLUSIONS@#The 16 X-STR loci meet the application requires of forensic genetics, especially for testing the special paternity cases.

Establishment and Verification of 6-color Fluorescent-labeled Rapid PCR Amplification System / 法医学杂志

OBJECTIVE@#To establish the rapid PCR amplification program and system and to verify the technical indexes.@*METHODS@#PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed.@*RESULTS@#The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate.@*CONCLUSION@#The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.

Application of Multiple Genetic Markers in a Case of Determination of Half Sibling / 法医学杂志

OBJECTIVE@#A case of half sibling was determined with multiple genetic markers, which could be potentially applied for determination of half sibling relationship from same father.@*METHODS@#Half sibling relationship was detected by 39 autosomal STR genetic markers, 23 Y-chromosomal STR genetic markers and 12 X -chromosomal STR genetic markers among ZHAO -1, ZHAO -2, ZHAO -3, ZHAO -4, and ZHAO-5.@*RESULTS@#According to autosomal STR, Y-STR and X-STR genotyping results, it was determined that ZHAO-4 (alleged half sibling) was unrelated with ZHAO-1 and ZHAO-2; however, ZHAO-3 (alleged half sibling), ZHAO-5 (alleged half sibling) shared same genetic profile with ZHAO-1, and ZHAO-2 from same father.@*CONCLUSION@#It is reliable to use multiple genetic markers and family gene reconstruction to determine half sibling relationship from same father, but it is difficult to determination by calculating half sibling index with ITO and discriminant functions.

A population genetic study of 22 autosomal loci of single nucleotide polymorphisms / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate polymorphisms and forensic efficiency of 22 non-binary single nucleotide polymorphism (SNP) loci.</p><p><b>METHODS</b>One hundred ethnic Han Chinese individuals were recruited from Dongguan, Guangdong. The 22 loci were genotyped with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS).</p><p><b>RESULTS</b>Nine loci were found with a single allele, 4 loci were found to be biallelic, whilst 9 loci were found to have 3 alleles. For 13 polymorphic loci, the combined discrimination power and power of exclusion were 0.999 98 and 0.9330, respectively. For the 9 non-biallelic loci, the combined discrimination power and power of exclusion were 0.9998 and 0.8956, respectively. For motherless cases, the combined power of exclusion was 0.6405 for 13 polymorphic SNPs and 0.6405 for 9 non-binary SNPs.</p><p><b>CONCLUSION</b>Non-binary loci have a greater discrimination power and exclusion power per SNP.</p>

Genetic polymorphism of 6 short tandem repeat loci in Chinese Han population in Shanxi / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To obtain the polymorphism data of six short tandem repeat (STR) loci, i.e. D9S925, D11S2368, D14S608, D15S659, D17S1290 and D20S470, in Chinese Han population in Shanxi province, and to evaluate the usefulness of the polymorphism data in forensic science.</p><p><b>METHODS</b>The D9S925, D11S2368, D14S608, D17S1290 and D20S470 primers were synthesized according to GenBank, while the D15S659 primers were self-designed. These primers were used to amplify DNA from 194 unrelated individuals collected from Shanxi Han population. Then the amplified fragments were separated by electrophoresis in 3130 Genetic Analyzer. GeneMapper3.2 software was used to analyze the results.</p><p><b>RESULTS</b>The distributions of genotypes for the six STR loci were in accordance with Hardy-Weinberg equilibrium. The polymorphism information content (PIC) of the six STR loci were from 0.750 to 0.860, and the heterozygosity ranged from 0.756 to 0.894, and the discrimination power were from 0.920 to 0.965, and the probability of exclusion were from 0.519 to 0.784. The combined probability of exclusion and the combined discrimination power were 0.9988 and 0.99999998, respectively.</p><p><b>CONCLUSION</b>The data indicated that D9S925, D11S2368, D14S608, D15S659, D17S1290 and D20S470 loci have high probability of exclusion and discrimination power. They can be used as genetic markers in forensic test and personal identification.</p>

The Y-STR polymorphisms and phylogenetic relationships of two minority populations in Liaoning province / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the genetic polymorphisms of 11 Y-chromosomal short tandem repeats (Y-STR) loci in 484 male individuals from two minority populations, the Hui and Xibe, of Liaoning province, and to evaluate their forensic application values and genetic relationships with other 15 populations of China.</p><p><b>METHODS</b>Eleven Y-STR loci in all samples were amplified with PowerPlex Y System, and the PCR products were analyzed by 310 Genetic Analyzer. Cluster analysis and neighbor-joining tree were applied to show the genetic distance among the populations.</p><p><b>RESULTS</b>In Hui people, 187 haplotypes were identified, and the overall haplotype diversity value was 0.9990. The gene diversity values (GD) for each locus ranged from 0.4783(DYS437) to 0.9679(DYS385a/b); In Xibe people, 237 haplotypes were identified, and the overall haplotype diversity value was 0.9984. The GD value for each locus ranged from 0.3618(DYS391) to 0.9686(DYS385a/b). Comparing with 15 reference populations, the genetic distance between the Hui and Xibe was the nearest (0.0257), and that between the Hui and Yi was the farthest (0.1046), while the genetic distance between Xibe and Korean was also the farthest (0.0978). The NJ tree was similar to the results of clustering analysis and all the 17 populations were clustered into 3 groups.</p><p><b>CONCLUSION</b>The genetic distribution of the 11 Y-STR loci in Liaoning Hui and Xibe ethnic groups showed favorable polymorphisms, therefore are suitable for forensic identification and paternity testing in the local area. The study of haplotype diversity among different populations is useful in understanding their origins, migrations and their relationships.</p>

Polymorphisms of 11 Y-chromosomal short tandem repeat loci in Chongqing Tujia ethnic group and genetic relationships with 16 populations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the genetic polymorphisms of 11 Y-chromosomal short tandem repeats (STR) loci in Chongqing Tujia population, and to evaluate their forensic application values and genetic relationships with the other 16 populations of China.</p><p><b>METHODS</b>Eleven Y-STR loci in 215 unrelated Tujia individuals from Chongqing were amplified with PowerPlex Y System, and the PCR products were analyzed by 310 Genetic Analyzer. Cluster analysis and phylogenic trees were applied to show the genetic distance among the populations.</p><p><b>RESULTS</b>A total of 195 haplotypes were identified and the overall haplotypes diversity for the 11 Y-STR loci was 0.9942. The gene diversity values (GD) for each locus ranged from 0.3757 (DYS391) to 0.9170 (DYS385a/b). Comparing with other 16 populations, the genetic distance between Tujia and Tibetan was the nearest (0.02467), that between the Tujia and Korean ethnic groups was the farthest (0.25350).</p><p><b>CONCLUSION</b>The genetic distribution of the 11 Y-STR loci in Chongqing Tujia population showed favorable polymorphisms. They are suitable for forensic identification and paternity testing in the local area. The study of genetic diversity among different populations is useful in understanding their origins, migrations and their relationships.</p>

Twenty Y-STR locus multiplex amplification and genetic polymorphism analysis in Han population in Chaoshan area / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study genetic polymorphisms of 20 Y-chromosomal short tandem repeats (STR) in Chaoshan Han population, and to evaluate their value in forensic science.</p><p><b>METHODS</b>Twenty Y-specific STR loci (DYS434, Y-GATA-A10, Y-GATA-H4, DYS438, DYS439, DYS443, DYS444, DYS446, DYS447, DYS448, DYS456, DYS458, DYS460, DYS520, DYS531, DYS557, DYS622, DYS630, DYS635 and DYS709) were amplified by using three fluorescence-labeled multiplex PCR systems and were analyzed by ABI310 genetic analyzer. One hundred and fifty-eight unrelated male individuals of Han population in Chaoshan area were investigated to determine the distribution of allele frequencies and haplotype.</p><p><b>RESULTS</b>The Y-STR multiplexes developed had followed the published nomenclature and ISFG guidelines for STR analysis. Gene diversity ranged from 0.2506 at DYS434 to 0.8034 at DYS447. A total of 157 different haplotypes were observed, and among these, 156 were unique, while 1 was found for two times. The haplotype diversity value calculated from all 20 loci combined was 0.999998. None of Y-STR allele mutation was observed in the 30 father/ son pairs confirmed by autosomal STR analysis.</p><p><b>CONCLUSION</b>The results indicate that the 20 Y-STR loci are highly polymorphic and fathership genealogy inheritance are stable. The three fluorescence-labeled multiplex amplification systems that we constructed are suitable for forensic individual identification and paternity testing in Chaoshan area.</p>

Forensic application of 9 Y-STRs fluorescence-labeled multiplex amplification system / 法医学杂志

OBJECTIVE@#In order to increase significantly the discriminatory potential of Y-STR systems available to the forensic community, we have developed a system capable of simultaneously amplifying 9 Y-STR loci by fluorescence-labeled multiplex PCR technique.@*METHODS@#Primers of STR loci DYS434, GATA-A10, DYS438 and DYS439 were labeled with 6-FAM, primers of STR loci DYS531, DYS557, DYS448 were labeled with HEX, and primers of STR loci DYS456, DYS444 were labeled with TAMRA, respectively. PCR products were analyzed using capillary electrophoresis and GeneScan Software on the ABI Prism310 DNA Analyzer. Series experiments were carried out to evaluate the useful value in forensic application such as the sensitivity, male specificity and genotyping DNA different tissues of the same individual.@*RESULTS@#9 Y-STR loci were exactly determined following optimization of the polymerase chain reaction. In a sample of 120 males, a total of 105 different haplotypes was identified, 97 of them being unique. Overall, haplotype diversity was 0.996 8. It was proved that genotyping of these 9 Y-STR loci in some sexual crime should be prior to that of automosomal STR.@*CONCLUSION@#The results suggest that the newly constructed 9-plex will be very powerful for establishing Y-STR database, population genetic studies and mixture stains identification.

DNA samples preparation from single cell and its application in sensitivity test / 法医学杂志

OBJECTIVE@#To establish a reliable, exact and practical method to prepare DNA samples for sensitivity-test purposes.@*METHODS@#The micromanipulation method was employed to prepare exact quantity DNA samples used to study the sensitivity of Profiler Plus Kit-ABI PRISM 310 system.@*RESULTS@#We succeed in establishing a micromanipulation method to prepare groups of DNA samples, which contain 1-11 cells in turn, and also succeed in using them to study the sensitivity of Profiler Plus Kit-ABI310 system.@*CONCLUSION@#The method we have established is proved to be a reliable, exact and practical way to prepare DNA samples for sensitivity-test purposes.

Distribution of haplotypes for four Y-sTR loci and validation in forensic science by using a double-fluorescent multiplex PCR system / 法医学杂志

OBJECTIVE@#We focus on developing a multiplex PCR system for Y-STR loci that can be detected by double fluorescent system and assessing their usefulness in forensic mixture samples.@*METHODS@#The primers of four Y-STR loci (DYS-GATA-A10, DYS531, DYS557 and DYS448) amplified by multiplex PCR technique were labeled with fluorescence, then the PCR products of these Y-STRs loci were detecting and typing by ABI PRISM310 Genetic Analyzer.@*RESULTS@#When 120 unrelated individuals from the Han population in Chengdu were detected by the system, Y-GATA-A10, DYS531, DYS557 and DYS448 showed 5, 5, 8, 7 alleles, respectively. A total of 78 different haplotypes was identified and the genetic diversity reached 0.9881. To the three cases of mixture stains failed by using conventional autosomal STR analysis, our multiplex system drew conforming conclusion comparing to the suspect's Y-STRs genotypes.@*CONCLUSION@#Our results show that the multiplex system of four Y-STR will be very powerful for Y-STR database establishing, the paternity testing and mixture stains identifying.

Constructing standard allelic ladders for four short tandem repeat loci and employing them in a population study on Han Nationality of Chengdu in China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To solve the problems in the accuracy and standardization of short tandem repeats-polymerase chain reaction (STR-PCR) typing, the authors adopted the molecular clone technology in producing the standard allelic ladders of D1S1676, D2S2735, D11S1977 and D22S444 loci and applied them in a population study on the Hans in Chengdu, China.</p><p><b>METHODS</b>PCR was used to produce several different allelic fragments of these loci. PCR products were eluted from the gel and re-amplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pGEMR-T plasmid vectors and the recombinant were transfected into competent E.coli DH5alpha TM cells. The results of sequencing confirmed that the size and the construction of the inserts were correct. The recombinant plasmids DNA with the inserts were then used as template for re-amplification to generate the four loci standard ladders.</p><p><b>RESULTS</b>The authors succeeded in producing large quantity of standard allelic ladder of these four loci, with which the genetic polymorphisms of these loci in Chengdu Han population of China were studied.</p><p><b>CONCLUSION</b>This method is of high value for forensic DNA typing to construct standard ladders. D1S1676, D2S2735 loci are robust for forensic analysis in Chinese Han population, whereas the value of D11S1977 and D22S444 loci is limited.</p>

Genetic polymorphisms of four STR loci on chromosome X and their forensic applications in a Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To add DXS7133, GATA198A10, DXS9896 and DXS6797 to the panel of forensically validated X chromosome markers, and apply the multiplex amplification system to a population study and forensic analysis on the Hans of Chengdu.</p><p><b>METHODS</b>The PCR products were detected by the polyacrylamide gel electrophoresis and silver staining method. Hardy-Weinberg equilibrium of females was tested and every forensically interested value was calculated.</p><p><b>RESULTS</b>Sequencing revealed that their common sequence motifs were tetranucleotide repeats. Population genetic data were obtained by analyzing 120 unrelated females and 100 males from Chengdu Han ethnic group. In this population, DXS7133, GATA198A10, DXS9896 and DXS6797 exhibited 6, 6, 11, 8 distinguishable alleles respectively. Chi-square test demonstrated that genotype frequencies in females did not depart from Hardy-Weinberg equilibrium. Power of discrimination for female samples for the four loci were 0.7962, 0.8021, 0.9675, and 0.9444. The parentage testing in 32 cases revealed a typical X-linked inheritance and no mutations.</p><p><b>CONCLUSION</b>DXS7133, GATA198A10, DXS9896 and DXS6797, which are highly polymorphic in Chengdu Han population, are appropriate for individual identification and paternity testing involving a female child.</p>