Two dinucleotide repeat polymorphisms at the D8S1218 and D8S1219 loci

Summary Two polymorphic dinucleotide (CA) repeat dones were isolated from a CEPH mega-YAC clone (844E2), and were localized to chromosome 8 using a panel of 13 mouse/human somatic cell hybrids.     

Two dinucleotide repeat polymorphisms at the D8S1444 and D8S1445 loci

Summary Two polymorphic dinucleotide (CA) repeat clones were isolated from two P1 phage clones, 1239F3 and 8H11, and were localized to chromosome 8 using a panel of 13 mouse human somatic cell hybrids.     

Two dinucleotide repeat polymorphisms at the D8S1442 and D8S1443 loci

Two polymorphic dinucleotide (CA) repeat clones were isolated from cosmids, cCI8-1121 and cCI8-1199, mapped to chromosome 8p11.2-p12.     

广西黑衣壮族D2S1338,D8S1179,D18S51基因座遗传多态性

目的 获得广西黑衣壮族人群3个STR基因座的群体遗传分布资料.方法 应用荧光标记STR基因扫描技术对152名广西黑衣壮族无关个体进行基因分型.结果 D2S1338检测出11种等位基因,频率分布在0.0030～0.2800.D8S1179检测出7种等位基因,频率分布在0.0987～0.2237.D18S51检测出12种等位基因,频率分布在0.0033～0.2467.对上述3个STR基因座的基因型观察值和期望值进行X^2检验,均符合Hardy-Weinberg平衡定律（P＞0.05）.D2S1338、D8S1179、D18S51基因多样性分别为0.8498、0.8450、0.8507;多态信息总量均为0.8300.结论 广西黑衣壮族3个STR基因座属高度多态性位点;D2S1338的等位基因19、D8S1179的等位基因10和D18S51的等位基因15可能是广西黑衣壮族群体中相应STR基因座最原始的等位基因.     

D6S296、D8S264基因座多态性与广西地区汉/壮族人群精神分裂症的关联性

目的探讨D6S296、D8S264基因座多态性与广西地区汉、壮族人群精神分裂症的关联性以及这两个基因座与汉、壮族精神分裂症关联性的差异。方法选取广西脑科医院汉族精神分裂症患者及其健康直系家属46对(其中患者46例,家属49例),壮族精神分裂症患者及其健康直系家属45对(其中患者45例,家属48例)作为研究对象,分别以其健康家属作为正常对照组。提取其全血DNA并使用PCR扩增仪对其进行扩增,所得PCR产物用ABI3730XL型基因分析仪自动进行基因检测,并用遗传学统计方法进行分析。结果汉、壮族正常对照组D6S296、D8S264各等位基因频率的分布均符合Hardy-Weinberg遗传平衡定律(P0.05)。汉、壮族精神分裂症患者组比较,D6S296的264bp等位基因频率分别为2.2%和18.9%(2/17),差异有显著统计学意义(χ2=13.6,P0.01);D8S264的129bp等位基因频率分别为18.5%/8.0%(17/7),差异有统计学意义(χ2=4.31,P0.05),其他各等位基因频率的分布在各组间的差异均无统计学意义(P0.05)。结论广西地区汉、壮族人群精神分裂症的发生可能与D6S296、D8S264多态性无关。     

成都汉族群体七个短串联重复序列基因座的遗传多态性和法医学应用研究

目的获得中国成都地区汉族群体DIS2142、DIS3733、D2S1774、D3S2459、D21S1409、D21S1437和D21S2055七个短串联重复序列（shoa tandem repeam，STR）基因座的群体遗传学资料，评价它们在法医鉴定的应用价值。方法用PCR、聚丙烯酰胺凝胶电泳、银染技术，对283名成都汉族无血缘关系的个体及50个家庭样品进行检测。结果DIS2142检出11个等位基因，23种基因型；DIS3733检出8个等位基因，19种基因型；D2S1774检出8个等位基因，15种基因型；D3S2459检出7个等位基因，19种基因型；D21S1409检出6个等位基因，12种基因型；D21S1437检出9个等位基因，26种基因型；D21S2055检出20个等位基因，77种基因型；基因型分布符合Hardy．Weinberg平衡定律；50个家庭样品调查证实上述基因座均符合孟德尔常染色体共显性遗传，未发现突变。结论DIS2142、D1S3733、D2S1774、D3S2459、D21S1409、D21S1437和21S2055基因座具有较好的多态性。基因座间独立性分析，证实上述7个基因座之间不存在连锁关系，可作为法医学亲子鉴定和个人识别的遗传标记。     

青岛地区D6S477等五个短串联重复序列基因座的遗传多态性研究

目的 了解D6S477等5个基因座在青岛地区汉族群体中基因型分布及等位基因频率等遗传多态性数据，初步探讨其应用价值。方法 收集200名青岛地区汉族无关个体的静脉血，ACD抗凝，采用Chelex法提取DNA，应用聚合酶链反应技术，扩增D6S477、D9S1118、D18S865、D19S400和D20S161基因座的短串联重复序列，聚丙烯酰胺凝胶垂直电泳，银染显色分型。结果 获得了青岛地区汉族群体上述5个基因座的等位基因频率，基因型分布均符合Hardy-Weinberg平衡（P〉0．05）。结论 这5个基因座在青岛地区汉族群体中有较高的非父排除率和个人识别机率，在遗传学研究中有较高的应用价值。     

Genetic variation at 5 new autosomal short tandem repeat markers (D10S1248, D22S1045, D2S441, D1S1656, D12S391) in a population-based sample from Maghreb region

Aim

To investigate allele distribution and genetic parameters of a population-based sample from Maghreb region.

Methods

Allele frequencies for 5 new autosomal short tandem repeat (STR) markers (D10S1248, D22S1045, D2S441, D1S1656, and D12S391) and several forensic parameters were determined for 95 unrelated individuals.

Results

The combined power of discrimination and power of exclusion for the 5 loci were high (0.9999991 and 0.9954757, respectively). Allele frequencies were compared with previously published population data. Significant differences were found between Maghreb population and all other populations at the locus D2S441. Also, significant differences were found between the Maghreb and the African American population at the D22S1045, D1S1656, and D12S391 loci, between Maghreb and Caucasian population at the D1S1656 locus, and between Maghreb and Hispanic population at the D22S1045 locus.

Conclusions

Typing of the 5 new STR loci may provide a useful addition to the previously established sets of autosomal STRs.Short tandem repeats (STR) are widely used for forensic testing. Ordinary paternity cases are solved by commercially available multiplexes kits, however, for more difficult cases, such as complex kinship analysis, additional STRs are needed to obtain better results. Besides, as many national DNA databases are growing and a large number of comparisons are being made within and between databases, concern for possible false-positive results may arise. This increases the need to introduce additional loci. The first European Standard Set (ESS) of loci included only 7 STRs loci, but the European Network of Forensic Science Institutes and the European DNA Profiling recommended to extend the ESS loci by adopting additional 3 miniSTRs loci (D10S1248, D22S1045, D2S441) and 2 additional polymorphic loci in 2006 (D1S1656, D12S391) (1,2).These new 5 loci improve the discriminatory power of forensic analysis and, by amplifying fragments well below current average amplicon sizes, can enhance genotyping success when analyzing highly degraded DNA (3,4).In order to verify and allow their use in forensics, the usefulness of ESS STR loci, it is necessary to obtain sufficient data from different populations.     

Allele-specific MVR-PCR analysis at minisatellite D1S8

Minisatelilte variant repeat mapping by the polymerase chainreaction (MVR-PCR) provides a digital approach to DNA typingof great potential use both in forensic medicine and, by mappingsingle alleles, for exploring allelic variability and mutationprocesses at minisateliltes. The MVR haplotypes of single allelescan be determined either from physically separated alleles orby pedigree analysis of digital diplold codes generated fromboth alleles simultaneously. We now show that single allelescan be rapidly mapped from total genomic DNA using allele-specificPCR primers directed to polymorphic sites in the DNA flankingthe minisatellite. This approach can also be used to dissectmixed DNA samples such as those often encountered in forensicDNA analysis.     

Genetic parameters of five new European Standard Set STR loci (D10S1248, D22S1045, D2S441, D1S1656, D12S391) in the population of eastern Croatia

Aim

To establish allele frequencies and genetic parameters in eastern Croatia population and to compare them with those in other populations. The second aim was to compare the genetic profiles obtained with different forensic kits amplifying the same genetic markers.

Methods

Blood samples of 217 unrelated individuals from eastern Croatia were genotyped using AmpFlSTR NGM kit. Allele distribution and other genetic parameters were determined for 15 short tandem repeat (STR) loci, including the 5 loci recently added to the European Standard Set (ESS) of STR loci (D10S1248, D22S1045, D2S441, D1S1656, and D12S391). Ninety-six samples underwent duplicate analysis using AmpFlSTR Identifiler kit.

Results

Power of discrimination was highest for the two new ESS loci, D1S1656 (0.97254) and D12S391 (0.97339). Comparison of allele frequencies for 5 new ESS loci in our sample with previously published population data showed a significant difference from Maghreb population on D2S441 and from American Caucasian population on D1S1656. Comparison of allele frequencies for standard 10 STR loci with all the neighboring populations’ data showed a significant difference only from Albanian population (on D2S1338, D18S51, and TH01). Discordant genotypes were observed in 5 (5.2%) samples at a single locus when amplified with both AmpFlSTR NGM and AmpFlSTR Identifiler kit.

Conclusion

New ESS STR loci are highly polymorphic and short, and therefore very useful for the analysis of challenging forensic samples. DNA samples purposed for establishing databases should be routinely amplified in duplicate.To facilitate DNA profiles comparison between databases of different European countries, The European Network of Forensic Science Institutes (ENFSI) and European DNA Profiling Group (EDNAP) have recently added five new loci (D10S1248, D22S1045, D2S441, D1S1656, and D12S391) to the European Standard Set of short tandem repeat (STR) loci (1,2). These new loci were included into the AmpFlSTR NGM PCR amplification kit (NGM kit; Life Technologies, Foster City, CA, USA).The Laboratory for DNA Analysis in Osijek was established to participate in the identification of missing persons after the war in Croatia (1991-1995). In collaboration with the laboratories in Zagreb and Split, a database of genotypes of missing persons’ relatives was created including approximately 5000 persons. The greatest part of the included genetic information is based on the 15 loci incorporated in AmpFlSTR Identifiler PCR amplification kit (Identifiler kit; Life Technologies, Foster City, CA, USA). Skeletal remains are identified by comparing the genotype of each piece of skeletal remains with the genotypes in the missing persons’ relatives database. Such non-targeted matching in a database containing several thousands genotypes considerably decreases the reliability of the established match. Still, the majority of identified skeletal remains were matched in such a way, as genotypes of the missing persons from the father-mother-child trio. Even within so large a database, hundreds of genotypes of skeletal remains still do not have a match, due to a lack of adequate relatives. Matching a profile created from a piece of skeletal remains across the whole database returns many adventitious matches, partly because some genotyped loci have low discrimination power (2). An especially large number of adventitious matches is present if the genetic profile from skeletal remains is partial.In a targeted approach to DNA typing, loci on the Y-chromosome and mtDNA can be amplified, but at a database level more useful are the loci on somatic chromosomes. Evidential value of a genetic match based on STR typing relies on high polymorphism and a large number of STR loci. In order to obtain as much as possible genetic information, we used the NGM kit. Our aim was to increase the number of genetic markers in order to achieve higher evidential value of STR typing and to amplify short STR loci, often better preserved in degraded samples. Especially valuable are three new “mini” STR loci (D10S1248, D22S1045, and D2S441), engineered to produce short amplicons (up to 150 bp) that are more successfully obtained from the most degraded samples. The remaining two new loci (D1S1656 and D12S391) are also relatively short and highly polymorphic (3-7). Besides obtaining information on the 5 new loci, the NGM kit includes improved chemistry that maximizes performance on challenging samples.In the new European Standard Set (ESS) of STR loci, allele distribution and genetic parameters still have to be determined. A population study on the new loci has been performed for several countries (including Belgium, Germany, Hungary, Maghreb countries, Poland, and USA) (7-12). However, there has been no such study either for Croatian or its neighboring populations. Therefore, we carried out a population study on a sample from eastern Croatia, which might be the most appropriate regional sample, because this part of the country sustained the greatest human losses during the war. Since the greatest part of our relatives’ database is based on the Identifiler kit, which shares 10 loci with NGM kit (D3S1358, vWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01, and FGA), we compared the genetic profiles obtained with both of these kits amplifying the same genetic markers. We also compared the obtained genetic parameters for 15 STR loci with the available population data from the neighboring countries.     

Dinucleotide repeat polymorphism at the D8S1223 locus

Summary A polymorphic dinucleotide (CA) repeat clone was isolated from a CEPH mega-YAC clone (936F7), and was localized to chromosome 8 using a panel of 13 mouse/human somatic cell hybrids.     

微卫星D8S84和D8S85在家族性热性惊厥人群相关性分析中的应用

目的：确认家族性热性惊厥与８号染色体长臂（８ｑ１３－２１）的关联；方法：用微卫星二核苷酸重复序列Ｄ８Ｓ８４和Ｄ８Ｓ８５对１３５例正常人和６３个热性惊厥家系进行聚合酶链反应－变性聚丙烯酰胺凝胶电泳（ＰＣＲ－ＰＡＧＥ）分型，结果用遗传学群体与家系计算机系统ＰＰＡＰ统计；结果：两种标志基因在中国正常人群处于Ｈａｒｄｙ－Ｗｅｉｎｂｕｒｇ平衡，并有较西方人略高的多态信息含量（ＰＩＣ），Ｄ８Ｓ８５的两种单体型频率在热性惊厥群体和普通正常人群存在显著性差异；结论：家族性热性惊厥可能与８ｑ的某些位点相关     

广东汉族人群21号染色体4个短串联重复序列多态性

目的调查广东地区汉族人群4个短串联重复序列D21S1433、D21S1442、D21S1444、D21S2051的多态性分布。方法采用荧光定量PCR（quantitative fluorescence PCR，QF-PCR）技术，对广东地区200份无亲缘关系样本进行等位基因分型，计算4个短串联重复序列的基因频率、基因型频率，等位基因杂合度，期望杂合性、多态信息含量和非父排除率。结果D21S1433、D21S1442、D21S1444、D21S2051分别检出9、10、9、5个等位基因，等位基因型分布均符合Hardy-Weinberg平衡（P〉0．05）。D21S1433和D21S1442的杂合度较高，分别为0．818和0．820，D21S2051的杂合度最低，为0.261。结论D21S1433、D21S1442、D21S1444具有较高的杂合度和多态信息量，作为遗传标记较为理想；D21S2051杂合度较低，不能作为遗传标记物。     

吉林朝鲜族D4S2368、D6S1043和D9S925基因座位的遗传多态性

目的了解吉林延边地区朝鲜族人群3个短串联重复序列(short tandem repeats,STR)基因座D4S2368、D6S1043、D9S925的遗传多态性分布,获取相应多态位点的群体遗传学数据。方法采用PCR扩增技术及聚丙烯酰胺凝胶电泳方法,对310名朝鲜族个体的3个STR基因座的等位基因频率进行分析。结果在3个基因座D4S2368、D6S1043、D9S925分别检出7、13、9个等位基因,除D6S1043(P=0.011)以外,其他两个基因座多态性分布均符合Hardy-Weinberg平衡定律(P>0.05)。杂合度大于0.717,多态信息含量大于0.670,累积个人识别力及非父排除率分别为0.9995和0.952。结论上述3个基因座均具有较高的杂合度和多态信息量,是较理想的遗传标记系统,对朝鲜族群体的亲子鉴定、个人识别及民族基因库的建立具有重要意义。     

内蒙古农区蒙古族D3S1358、D13S317和D5S818 基因座位的遗传多态性

目的　了解内蒙古蒙古农区族人群 3个短串联重复序列 ( shorttandem repeats,STR)基因座 D3S135 8、D13S317、D5 S818的遗传多态性分布 ,获取相应多态位点的群体遗传学数据。方法　采用PCR扩增技术及聚丙烯酰胺凝胶电泳方法 ,对 2 91名蒙古族个体的 3个 STR基因座的等位基因频率进行分析。结果　 3个基因座 D3S135 8、D13S317、D5 S818分别检出 6、10、8个等位基因 ,多态性分布均符合Hardy- Weinberg平衡定律 ( P>0 .0 5 )。杂合度大于 0 .7332 ,多态信息含量大于 0 .6 884 ,累积个人识别力及非父排除率分别为 0 .9991和 0 .980 6。结论　上述 3个基因座均具有较高的杂合度和多态信息量 ,是较理想的遗传标记系统 ,对建立蒙古族民族基因库和进一步研究群体的遗传变异具有重要意义     

广西黑衣壮族D6S477、D18S865、D19S400、D20S161的遗传多态性

目的:调查广西黑衣壮族人群4个短串联重复序列(short tandem repeats,STR)基因座:D6S477、D18S865、D19S400、D20S161的群体遗传分布资料。方法:应用PCR扩增结合聚丙烯酰胺凝胶电泳技术分析180名广西黑衣壮族无关个体。结果:4个STR基因座平均观察杂合度分别为0.8361;平均多态信息总量为0.8047;累积个体识别力达0.999 988 391,累积非父排除率达0.988 207。在D19S400、D20S161基因座上广西黑衣壮族与广西武鸣壮族、广州汉族与青岛汉族的等位基因频率分布差异均无统计学意义;广西黑衣壮族与广西武鸣壮族遗传距离为0.0304。结论:D6S477的等位基因15、D18S865的等位基因11、D19SA00的等位基因12、D20S161的等位基因18可能是广西黑衣壮族群体中相应STR基因座最原始的等位基因。广西黑衣壮族的4个STR基因座属高度遗传多态性标记。同一民族内不同支系的遗传关系近于不同民族间的遗传关系。     

广西壮族D6S477等四个STR基因座的遗传多态性

目的 研究广西壮族人群4个STR（short tandem repeats）基因座：D6S477、D18S865、D19S400、D20S161的遗传多态性。方法 应用PCR扩增结合聚丙烯酰胺凝胶电泳技术分析200名广西壮族无关个体。结果 4个STR基因座平均期望杂合度为0．8442，平均多态信息总量为0．8247，累积个体识别力达0．999994474，累积非父排除率达0．9856。D6S477的等位基因15、D18S865的等位基因10和11、D19S400的等位基因11、D20S161的等位基因18可能是广西壮族群体中相应STR基因座最原始的等位基因。结论 广西壮族的4个STR基因座属高度遗传多态性。     

D16S539、D7S820和D13S317位点在新疆哈萨克族中的遗传多态性

目的 了解新疆哈萨克族人群D16S539、D7S820和D13S317 3个STR位点的遗传多态性。方法 应用多重PCR扩增，6％变性聚丙烯酰胺凝胶电泳结合银染技术对102名无关个体及8个家系42人的哈萨克族人群进行调查，并与其他人群进行了比较。结果 3个位点分别检测出8、7、8个等位片段，多态性分布符合Hardy-Wdinberg平衡定律，期望杂合度为：0.9439、0.9356、0.9304，累积多态信息量为0.9905、个体识别为0.9998、非父排除率为0.9572，与其他人群比较差异有显著性（P＜0.05），在家系调查中无一突变发现，均按孟德尔遗传规律传递。结论 3个STR位点的综合检验在法医学应用及群体遗传学中显示了较高的价值。     

High density deletion mapping of bladder cancer localizes the putative tumor suppressor gene between loci D8S504 and D8S264 at chromosome 8p23.3

Deletion of chromosome 8p is associated with the progression of bladder cancer. To identify the putative tumor suppressor gene locus we have analyzed 145 bladder cancers with 12 microsatellite markers for allelic changes at the chromosome 8p23.3 region. We mapped the smallest overlapping deletion to approximately 0.7 cM genetic distance between loci D8S504 and D8S264. Allelic changes at this region occurred in 75 (52%) of the 145 tumors. We found a significant correlation between alterations at chromosome 8p23.3 and the tumor grade. The correlation between genetic changes and tumor stage reflected the distribution of tumors of different grades in each pathologic stage.