Y染色体特异STR位点应用于无创伤性产前胎儿遗传信息的研究

目的 建立利用孕妇血清进行无创伤性产前胎儿分子遗传信息分析的方法。方法 采集53名11～36孕周的孕妇的血清,利用血清中胎儿DNA,采用”Y-PLEX 6”试剂盒,复合扩增DYS393、DYS19、DYS389 Ⅱ、DYS390、DYS391和DYS385等6个Y-STR位点,PCR产物经基因测序仪电泳检测,用相关软件分析Y-STR基因型。结果 ①29名分娩出生证实为男婴的孕妇,均检测出特异性Y-STR等位基因。检测的6个Y-STR位点,以DYS393位点的检出率最高(29/29);其次是DYS19位点,检出率为62.07％(18/29);再次是DYS390位点,检出率为34.48％(10/29);其余的DYS389 Ⅱ、DYS391和DYS385位点的检出率则较低。②24名分娩出生证实为女婴的孕妇,均未检测出特异性Y-STR等位基因。③根据DYS393位点是否检测出特异性等位基因,以及该基因波峰的高度和波峰面积值,鉴定胎儿性别的准确率达100％。④29名妊娠男婴的孕妇的血清样本,检测出的Y-STR等位基因与“丈夫”的相一致。结论 本研究建立的无创伤性Y-STR分子遗传分析方法,具有多态性丰富、灵敏度高、特异性强等特点,提高了无创伤性产前胎儿性别遗传鉴定的准确性,同时为解决妊娠男婴的亲子鉴定提供了理论依据,具有广泛的应用前景。     

Haplotypes of 20 Y-chromosomal STRs in a population sample from southeast China (Chaoshan area)

In this study, 20 Y-specific short tandem repeat (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(Y-GATA-C4), and DYS709) were analyzed in 158 unrelated healthy men from southeast China by three fluorescence-labeled multiplex polymerase chain reaction systems. The Y-STR multiplexes developed have followed the published nomenclature and International Society for Forensic Genetics (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 two times. The haplotype diversity value calculated from all 20 loci combined was 0.9997, which is informative. Furthermore, 80 father–son pairs, previously confirmed by autosomal STR analysis, were typed using the same 20 Y-STR loci, and four mutation events were identified at the Y-GATA-H4, DYS439, DYS456, and DYS458 loci, giving an average mutation rate of 0.25% per locus per generation (95% confidence interval 0.09–0.54). These results including the haplotype data at 20 Y-STR loci would enrich Chinese genetic informational resources and provide useful information in forensic practice. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

南方汉族无关男性群体17个Y—STR基因座单倍型遗传多态性的研究

目的研究中国南方汉族无关男性群体中DYS426等17个Y—STR基因座的遗传多态性，以应用于法医学鉴定。方法采用AmpFlSTRYfiler^TMPCR复合扩增试剂盒和ABIprism^TM3100基因测序仪荧光检测方法．检测119份无关男性个体血样．调查南方汉族的17个Y—STR基因座的单倍型频率，并计算单倍型遗传多样性。结果在119份无关男性样本中，共检出118种Y—STR单倍型，其中117种单倍型仅出现1次。另一单倍型15／12／23／28／16／15／13，13／13／10／11／20／14，12，14，10，18出现2次．单倍型多样性达0.9999。结论AmpFISTRYfiler复合扩增试剂盒所包含的17Y—STR基因座，在南方汉族无关男性个体中具有丰富的遗传多态性和较高的非父排除能力．在法医学个体识别和亲子鉴定中具有广泛应用价值。     

壮族非亲缘男性个体中9个短片段Y-STR基因座遗传多态性的研究

本研究探讨中国最大少数民族壮族Y染色体特异短串联重复序列（Y-STR）单倍型的遗传多态性。采用多重PCR扩增和ABIPrism^TM3100基因测序仪荧光检测方法，对85名壮族非亲缘男性个体的9个短片段长度Y—STR基因座，进行基因频率和单倍型频率的调查。结果表明：在85名非亲缘男性个体中，除DYS426基因座多态性较低，其余8个基因座的GD值的分布在0．4387—0．8129之间。9个Y—STR基因座的单倍型共有70种．单倍型多样性达0．9926。结论：壮族非亲缘男性个体Y—STR单倍型的遗传多态性丰富，与我们以往的南方汉族非亲缘男性群体遗传资料相比较，具有显著的差异。     

Unbiased Interpretation of Haplotypes at Duplicated Microsatellites

The Y-chromosome is a powerful tool for population geneticists to study human evolutionary history. Haploid and largely non-recombining, it should contain a simple record of past mutational events. However, this apparent simplicity is compromised by Y-linked duplicons, which make up ∼35% of this chromosome; 25% of these duplicons are large inverted repeats (palindromes). For microsatellites lying in these palindromes, two loci cannot be easily distinguished due to PCR co-amplification, and this order misspecification of alleles generates an additional variance component. Due to this ambiguity, population geneticists have traditionally used an arbitrary method to assign the alleles (shorter allele to locus 1, larger allele to locus 2). Here, we simulate these posterior estimate distributions under three different novel allele assignment priors and compare this with the original method. We use a sample of 33 human populations, typed for duplicated microsatellites lying within palindrome P8, to illustrate our approach. We show that both intra- and inter-population statistics can be dramatically affected by order misspecification. Surprisingly, matrices of pairwise F-statistics or distance estimates appear far less sensitive to order misspecification and remain relatively unchanged under the priors considered, suggesting that these microsatellites can be considered as useful markers for population genetic studies using an appropriate data treatment. Duplicated microsatellites represent an attractive source of information to investigate the extensive structural polymorphism observed among human Y chromosomes, as well as processes of intra-chromosomal gene conversion acting between duplicons.     

Temporal differentiation across a West-European Y-chromosomal cline: genealogy as a tool in human population genetics

The pattern of population genetic variation and allele frequencies within a species are unstable and are changing over time according to different evolutionary factors. For humans, it is possible to combine detailed patrilineal genealogical records with deep Y-chromosome (Y-chr) genotyping to disentangle signals of historical population genetic structures because of the exponential increase in genetic genealogical data. To test this approach, we studied the temporal pattern of the 'autochthonous' micro-geographical genetic structure in the region of Brabant in Belgium and the Netherlands (Northwest Europe). Genealogical data of 881 individuals from Northwest Europe were collected, from which 634 family trees showed a residence within Brabant for at least one generation. The Y-chr genetic variation of the 634 participants was investigated using 110 Y-SNPs and 38 Y-STRs and linked to particular locations within Brabant on specific time periods based on genealogical records. Significant temporal variation in the Y-chr distribution was detected through a north-south gradient in the frequencies distribution of sub-haplogroup R1b1b2a1 (R-U106), next to an opposite trend for R1b1b2a2g (R-U152). The gradient on R-U106 faded in time and even became totally invisible during the Industrial Revolution in the first half of the nineteenth century. Therefore, genealogical data for at least 200 years are required to study small-scale 'autochthonous' population structure in Western Europe.     

Differential Y-chromosome Anatolian Influences on the Greek and Cretan Neolithic

The earliest Neolithic sites of Europe are located in Crete and mainland Greece. A debate persists concerning whether these farmers originated in neighboring Anatolia and the role of maritime colonization. To address these issues 171 samples were collected from areas near three known early Neolithic settlements in Greece together with 193 samples from Crete. An analysis of Y-chromosome haplogroups determined that the samples from the Greek Neolithic sites showed strong affinity to Balkan data, while Crete shows affinity with central/Mediterranean Anatolia. Haplogroup J2b-M12 was frequent in Thessaly and Greek Macedonia while haplogroup J2a-M410 was scarce. Alternatively, Crete, like Anatolia showed a high frequency of J2a-M410 and a low frequency of J2b-M12. This dichotomy parallels archaeobotanical evidence, specifically that while bread wheat ( Triticum aestivum ) is known from Neolithic Anatolia, Crete and southern Italy; it is absent from earliest Neolithic Greece. The expansion time of YSTR variation for haplogroup E3b1a2-V13, in the Peloponnese was consistent with an indigenous Mesolithic presence. In turn, two distinctive haplogroups, J2a1h-M319 and J2a1b1-M92, have demographic properties consistent with Bronze Age expansions in Crete, arguably from NW/W Anatolia and Syro-Palestine, while a later mainland (Mycenaean) contribution to Crete is indicated by relative frequencies of V13.     

Evolution and migration history of the Chinese population inferred from Chinese Y-chromosome evidence

Y-chromosomes from 76 Chinese men covering 33 ethnical minorities throughout China as well as the Han majority were collected as genetic material for the study of Chinese nonrecombinant Y-chromosome (NRY) phylogeny. Of the accepted worldwide NRY haplogroups, three (haplogroups D, C, O) were significant in this sample, extending previous assessments of Chinese genetic diversity. Based on geographic, linguistic, and ethnohistorical information, the 33 Chinese ethnical minorities in our survey were divided into the following four subgroups: North, Tibet, West, and South. Inferred from the distribution of the newfound immediate ancestor lineage haplogroup O^*, which has M214 but not M175, we argue that the southern origin scenario of this most common Chinese Y haplogroup is not very likely. We tentatively propose a West/North-origin hypothesis, suggesting that haplogroup O originated in West/North China and mainly evolved in China and thence spread further throughout eastern Eurasia. The nested cladistic analysis revealed in detail a multilayered, multidirectional, and continuous history of ethnic admixture that has shaped the contemporary Chinese population. Our results give some new clues to the evolution and migration of the Chinese population and its subsequence moving about in this land, which are in accordance with the historical records.The first two authors contributed equally to this study.     

A multicenter investigation with interphase fluorescence in situ hybridization using X- and Y-chromosome probes

Twenty-six laboratories used X and Y chromosome probes and the same procedures to process and examine 15,600 metaphases and 49,400 interphases from Phaseolus vulgaris-leucoagglutinin (PHA)-stimulated lymphocytes. In Part I, each laboratory scored 50 metaphases and 200 interphases from a normal male and a normal female from its own practice. In Part II, each laboratory scored 50 metaphases and 200 interphases on slides prepared by a central laboratory from a normal male and a normal female and three mixtures of cells from the male and female. In Part III, each laboratory scored 50 metaphases (in samples of 5, 10, 15, and 20) and 100 interphases (in samples of 5, 10, 15, 20, and 50) on new, coded slides of the same specimens used in Part II. Metaphases from male specimens were scored as 98–99% XY with no XX cells, and 97–98% of interphases were scored as XY with 0.04% XX cells. Metaphases from female specimens were scored as 96–97% XX with 0.03% XY cells, and 94–96% of interphases were scored as XX with 0.05% XY cells. Considering the data as a model for any probe used with fluorescence in situ hybridization (FISH), a statistical approach assessing the impact of analytical sensitivity on the numbers of observations required to assay for potential mosaicisms and chimerisms is discussed. The workload associated with processing slides and scoring 50 metaphases and 200 interphases using FISH averaged 27.1 and 28.6 minutes, respectively. This study indicates that multiple laboratories can test/develop guidelines for the rapid, efficacious, and cost-effective integration of FISH into clinical service. Am. J. Med. Genet. 76:318–326, 1998. © 1998 Wiley-Liss, Inc.     

Influences of history, geography, and religion on genetic structure: the Maronites in Lebanon

Cultural expansions, including of religions, frequently leave genetic traces of differentiation and in-migration. These expansions may be driven by complex doctrinal differentiation, together with major population migrations and gene flow. The aim of this study was to explore the genetic signature of the establishment of religious communities in a region where some of the most influential religions originated, using the Y chromosome as an informative male-lineage marker. A total of 3139 samples were analyzed, including 647 Lebanese and Iranian samples newly genotyped for 28 binary markers and 19 short tandem repeats on the non-recombinant segment of the Y chromosome. Genetic organization was identified by geography and religion across Lebanon in the context of surrounding populations important in the expansions of the major sects of Lebanon, including Italy, Turkey, the Balkans, Syria, and Iran by employing principal component analysis, multidimensional scaling, and AMOVA. Timing of population differentiations was estimated using BATWING, in comparison with dates of historical religious events to determine if these differentiations could be caused by religious conversion, or rather, whether religious conversion was facilitated within already differentiated populations. Our analysis shows that the great religions in Lebanon were adopted within already distinguishable communities. Once religious affiliations were established, subsequent genetic signatures of the older differentiations were reinforced. Post-establishment differentiations are most plausibly explained by migrations of peoples seeking refuge to avoid the turmoil of major historical events.