用PCR扩增分析中国汉族人DIS80位点的遗传多态性

和扩增片段长度多态性技术分析人类的ＤＩＳ８０位点的ＤＮＡ多态性。检测了１０９名无关中国汉族人，发现了１８个等位基因，５０种基因型，杂合性为０．７７，个人鉴别力为０．９６。观察的基因型频率的分布符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ遗传平衡。检测的二例二代３口之家的基因型结果说明该位点属孟德尔式遗传。该位点个人鉴别力高，ＰＣＲ方法简便，灵敏，可在个人识别和亲子鉴定中发挥重要作用。     

用Amp－FLP技术研究中国汉族群体D4S95和DXS52位点的遗传多态性

为研究中国汉族群体Ｄ４Ｓ９５和ＤＸＳ５２位点的遗传多态性，应用改进的Ｄ４Ｓ９５和ＤＸＳ５２位点的扩增片段长度多态性（Ａｍｐ－ＦＬＰ）分析技术，检测了中国汉族个体共２２２名。结果分别为，（１）Ｄ４Ｓ９５位点：在１０８名无关个体中发现了７个等位基因（片段长度为９１０～１１５０ｂｐ），１８种基因型，杂合性为０．７６，个人鉴别力为０．８７１（２）ＤＸＳ５２位点：在１１４名无关个体中（男性７０人，女性４４人）发现了１４个等位基因（片段长度为６９５～２４００ｂｐ），在４４名女性个体中发现了２２种基因型，杂合性为０．７７．个人鉴别力为男性０．８９，女性０．９３１检测了两个家系两代３口之家的两位点的基因型，证明该两位点的基因按Ｍｅｎｄｅｌ定律遗传，该技术在法医科学鉴定中具有重要的应用价值。     

成都地区汉族人群中FABP2和F13A1的遗传多态性

目的了解成都地区汉族群体中短串联重复序列ＦＡＢＰ２和Ｆ１３Ａ１的遗传多态性，获得ＦＡＢＰ２和Ｆ１３Ａ１两个基因座的群体遗传学数据。方法应用ＰＣＲ扩增技术结合聚丙烯酰胺凝胶水平电泳分型方法分别调查ＦＡＢＰ２和Ｆ１３Ａ１基因座在成都地区汉族群体中的基因型分布。结果ＦＡＢＰ２基因座在成都地区汉族群体中存在５个等位基因，基因频率分别为０．５１３，０．０９６，０．３３０，０．０５２，０．００９。杂合性为６０．８７％，基因型分布符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡。Ｆ１３Ａ１基因座在成都地区汉族群体中存在５个等位基因，基因频率分别为０．１２８，０．０５３，０．２８８，０．５２７，０．００４。杂合性６６．３７％，基因型分布不符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡，样本数据不能代表总体数据。结论ＦＡＢＰ２基因座的检测分型可用于法医学亲子鉴定与个人识别，Ｆ１３Ａ１基因座则不宜用于法医学亲子鉴定与个人识别。     

5-羟色胺转运体基因VNTR位点与双相情感性障碍的关联分析

目的探讨５－羟色胺转运体基因第２内含子ＶＮＴＲ区域在中国人群中的群体遗传学背景，以及与双相情感障碍的关系。方法利用ＰＣＲ等方法在１７０个随机健康中国人中对５－羟色胺转运体基因第２内含子的ＶＮＴＰ多态性位点进行了群体遗传学研究，并就该位点与双相情感障碍的关系进行了关联分析。结果该位点在中国人群中的多态信息含量为０．１２，杂合度为１２％，且符合孟德尔遗传方式；其基因型分布、等位片段频率在中国汉族人群与高加索人种间存在着极显著性差异（Ｐ＜１０－８）；在与情感性障碍的关联分析中发现该多态性位点的等位片段１０与双相情感障碍女性群体存在着显著的关联（Ｐ＝０．０４３）。结论该结果表明：５－羟色胺转运体基因ＶＮＴＲ的位点多态性在中国人群与高加索人群间存在极显著的差异；同时，在男女性双相情感障碍群体中的差别可能暗示了该疾病的发病机理在性别上的差异。     

STR位点D19S253和D8S1179的多态性研究

目的研究ＳＴＲ位点Ｄ１９Ｓ２５３和Ｄ８Ｓ１１７９的多态性，为法医学应用提供基础数据。方法应用ＰＣＲ及ＰＡＧ电泳技术对武汉地区２００多名汉族无关个体进行了调查。结果两位点各检出９个等位基因，首次获得汉族人群频率分布。两位点基因型频率分布符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡。家系调查证实了等位基因的传递遵循孟德尔遗传规律，观察１００次减数分裂未发现突变基因。Ｄ１９Ｓ２５３和Ｄ８Ｓ１１７９位点的杂合度观察值分别为０．８０８９和０．８７１２，多态性信息含量分别为０．７７５４和０．８２５８。两位点联合ＰＤ值为０．９９６６，累积非父排除率为０．８７９１。结论表明这两个多态位点在法医学个人识别及亲子鉴定中是很有价值的遗传标记系统。     

中国广东人CYP2F1基因遗传多态性研究

目的检测广东地区正常人群和鼻咽癌患者中细胞色素P450酶系CYP2F1基因的多态性，并分析该基因遗传多态性与鼻咽癌易感性的关联。方法采用直接测序法检测40例鼻咽癌患者全血标本中CYP2F1基因全部10个外显子的多态性变化。对于等位基因频率较高的多态性位点，进一步采用错配聚合酶链反应．限制性片段长度多态性检测368例鼻咽癌患者和344名正常对照人群中该位点的等位基因频率。结果在40例鼻咽癌样本中，共检测到CYP2F1基因的35个单核苷酸多态性。其中，10个单核甘酸引起编码的氨基酸改变，1个移码突变，15～16bp之间插入C引起移码突变（15-16ins C），该等位基因频率为25％。但病例-对照分析却未能显示该位点突变与鼻咽癌易感的相关性（P〉0．05）。结论中国广东人的CYP2F1基因遗传多态性位点较多，但暂未发现与鼻咽癌的易感性关联的单一多态位点，多个多态性位点或不同基因多态性位点的协同互补作用可能才是鼻咽癌发生发展的关键影响因素。     

中国人D13S314位点STR的多态特征及其价值探讨

Ｄ１３Ｓ３１４位于Ｗｉｌｓｏｎ’ｓ病（ＷＤ）基因近端侧，其短串联重序列（ＳＴＲ）具有高度多态性，应用ＰＣＲ方法结合变性聚丙烯酰胺凝胶电泳可迅速加以检测。本研究检测了８０名正常中国人该位点的等位片段，探讨其多态性分布特征并与西方人进行比较，表明该痊点在中国人具有长度多态性，多态信息含量（ＰＩＣ值）为０．８９，有较高的应用价值。     

DXYS267位点的遗传多态性及伴性遗传单核苷酸变异的应用

目的 调查DXYS267基因座遗传多态性，研究该座位伴性遗传单核苷酸的特征及其法医学应用价值。方法 应用聚合酶链反应和聚丙烯酰胺凝胶电泳对DXYS267进行群体调查，并作Hardy-Wein-berg平衡检验，计算法医学群体遗传学参数。依据该基因座伴性遗传的单核苷酸变异，设计新的引物，选择性扩增Y染色体的DXYS267短串联重复序列。结果 在118名中国汉族无关个体中，共发现6个等位基因，基因型频率分布符合Hardy-Weinberg平衡，基因座杂合度为0．6706，个体识别率为0．8433，非父排除率为0．5957。新引物可选择性扩增Y．短串联重复（short tandem repeat，STR）序列，在184名男性无关个体共发现4个等位基因，单倍型多样性值为0．6372。结论 DXYS267属一类X、Y染色体序列同源的STR序列位点，是一个高度多态性的系统。同时利用该位点的伴性单核苷酸变异，选择性扩增Y染色体上的STR序列，用于亲权鉴定和个人识别，尤其是混合斑男性检材的分型和性别鉴定有实用价值。     

中国人D13S314位点STR的多态性特征及其价值探讨

D13S314位于Wilson’s病（WD）基因近端侧，其短串朕重复序列（STR）具有高度多态性，应用PCR方法结合变性聚丙烯酸胺凝胶电泳可迅速加以检测。本研究检测了8o名正常中国人该位点的等位片段，探讨其多态性分布特征并与西方人进行比较，表明该位点在中国人群中具有长度多态性，多态信息含量（PIC值）为o．89，有较高的应用价值。     

内皮素—1基因内含子4Taq I片段多态性的研究

根据EDN1第4内含子设计相应引物,建立检测EDN1第4内含子TaqI多态性PCR＋RFLP技术。利用该技术,扩增到具有特异性的PCR产物,长为358bp。经限制性核酸内切酶TaqI酶酶切检测中国汉族人群该片段多态性状况。实验显示中国汉人群中存在该位点的多态性。基因T1的频率为0．664;T2基因型频率为0．336;T1T1基因型频率为0．418;T1T2基因型频率为0．492;T2T2基因型频率为0．090。该位点可作为遗传标记探讨中国人群中与EDN1相关遗传病的关系。     

用PCR法分析中国汉族群体D17S30位点遗传多态性

用聚合酶链式反应对中国汉族群体１２２名无关个体Ｄ１７Ｓ３０位点扩增片段长度多态性进行分析研究，发现１０个等位基因，片段大小为１６８－７９８ｂｐ，基因频率为０．００８２－０．２５８２，杂合性为７８％。５５种可能基因型中发现了２７种，对基因型的观察值和期望值进行Ｘ＾２检验，符合Ｈａｒｄｙ－Ｗｅｉｂｅｒｇ平衡定律。     

中国延边朝鲜族人群Penta D、E基因座的遗传多态性

目的了解中国吉林延边地区朝鲜族人群Penta D和Penta E短串联重复序列基因座的遗传多态性分布，获得相应群体遗传学数据。方法应用PCR扩增片段长度多态性分析方法对100名无血缘关系的朝鲜族个体进行调查。结果在Penta D基因座，观察到8个等位基因及22种基因型；在Penta E基因座，观察到16个等位基因及35种基因型。结论两个基因座等位基因片段多态性分布均符合Hardy．Weinberg平衡定律，而且显示很高的个人识别能力。所得数据资料可为中国朝鲜族人群的法医学个人识别、亲子鉴定及遗传学研究提供依据。     

D1S80 polymorphism,including a new variant,in a population sample from Barcelona (Spain) using polymerase chain reaction

Allele and genotype frequencies for the D1S80 (MCT118) locus have been determined in a population sample from Barcelona (Spain) using the polymerase chain reaction (PCR) amplification and nonradioactive detection. In a total of 216 unrelated individuals, 24 alleles (23 common and 1 rare variant) and 67 genotypes (64 common and 3 variants) were observed. The 216 individuals came from 162 blood samples taken for paternity studies, 16 bloodstains from forensic cases, and 38 root hairs from normal individuals. The D1S80 locus demonstrated a heterozygosity of 0.7916, and a power of discrimination of 0.9731. The distribution of genotypes is in agreement with expected values according to the Hardy-Weinberg equilibrium. Additionally, the population from Barcelona differs, significantly, from the Finnish population and also, but with lower differences, from a U.S.A. Caucasian population. © 1993 Wiley-Liss, Inc.     

中国朝鲜族人群6个短串联重复序列位点的遗传多态性

目的 了解中国朝鲜族人群D16S539，D7S820，D13S317，CSF1PO，TPOX，TH01 6个短串联重复序列（short tandem repeat,STR)位点的遗传多态性分布，获得相应多态位点的群体遗传学数据。方法 应用PCR扩增片段长度多态性分析方法对100名无血缘关系的朝鲜族个体进行了调查。结果 D16S539基因座，观察到6个等位基因，18种基因型；D7S820基因座，观察到7个等位基因，22种基因型；D13S317基因座，观察到7个等位基因，23种基因型；CSF1PO基因座，观察到6个等位基因，16种基因型；TPOX基因座，观察到6个等位基因，11种基因型；THO1基因座，观察到5个等位基因，12种基因型。结论 6个位点等位片段多态性分布均符合Hardy-Weinberg平衡定律。且具有较高的杂合度，所得到的等位基因频率等数据可以为中国朝鲜族人群法医个体识别、亲子鉴定及遗传学研究提供依据。     

Histocompatibility Antigens in two American Indian Tribes of French Guiana

Two South American Indian populations were typed for HLA antigens. In each, the individuals typed were related and their genealogies were known. Determination of their genotypes was done; there seemed to be neither excess nor deficiency in homozygotes. The antigens observed, A2, A9, Aw19.2 (Aw30-Aw31), A28 for the first locus and B5, Bw15, Bw35, Bw40 for the second locus are in accordance with those previously described for other South American Indians. The two populations belong to the same primary linguistic family Tupi-Guarani and they live in the same geographic area, but there was no intertribal marriage until recently. Genetic drift can explain the differences observed.     

Distribution of KIR genes in the Croatian population

The KIR locus with genes involved in immune processes is among the most polymorphic and structurally diverse human loci. KIR genes encode activating and inhibitory receptors that differ in specificity for HLA class I ligands and signaling potential. These receptors are expressed principally by natural killer (NK) cells and subpopulations of T cells. This study represents the first report of the distribution of KIR genes, KIR genotypes and KIR/HLA pairs in 121 unrelated healthy Croatian individuals. Twenty-three different genotypes were observed in the Croatian population and all 16 KIR genes known to date were found. The most frequent KIR genotype was the AA genotype. All individuals had at least one inhibitory KIR/HLA pair with the majority of individuals with three inhibitory KIR/HLA pairs. The most frequent KIR/HLA pair was the KIR2DL3/C1 group. Our results demonstrated the similarity of the Croatian population’s KIR repertoire with other Caucasian populations reported so far.     

云南省白族六个基因座的遗传多态性调查

目的了解云南省白族６个短串联重复序列基因座遗传多态性分布。方法应用复合扩增技术对ＣＳＦ１ＰＯ、ＴＰＯＸ、ＴＨ０１、Ｆ１３Ａ０１、ＦＥＳＦＰＳ和ｖＷＡ等６个ＳＴＲ基因座进行分析,采用高分辨率的聚丙烯酰胺凝胶电泳分离、银染法显影技术,对云南白族上述６个ＳＴＲ基因座进行遗传多态性调查。结果ＣＳＦ１ＰＯ基因座,观察到７个等位基因,１９个基因型;ＴＰＯＸ基因座,观察到５个等位基因,１１个基因型;ＴＨ０１基因座,观察到６个等位基因,１６个基因型;Ｆ１３Ａ０１基因座,观察到６个等位基因,１６个基因型;ＦＥＳＦＰＳ基因座,观察到７个等位基因,１５个基因型;ｖＷＡ基因座,观察到７个等位基因,２１个基因型。结论上述６个短串联重复序列基因座基因频率分布与Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡吻合良好。     

Molecular analysis of the 18S rRNA gene of Cryptosporidium parasites from patients with or without human immunodeficiency virus infections living in Kenya,Malawi, Brazil,the United Kingdom,and Vietnam

An 840-bp fragment of the 18S rRNA gene was used to identify Cryptosporidium spp. recovered from human immunodeficiency virus (HIV)-infected and -uninfected patients from Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. Initial identification was by Ziehl-Neelsen acid-fast staining. Confirmation was by nested PCR, targeting the most polymorphic region of the 18S rRNA gene. Genotyping was by restriction endonuclease digestion of the PCR product followed by nucleotide sequencing. Among 63 isolates analyzed, four genotypes of Cryptosporidium were identified; 75% of the isolates were of the C. parvum human genotype, while the potentially zoonotic species were of the C. parvum bovine genotype (21.7%), the C. meleagridis genotype (1.6% [one isolate]), and the C. muris genotype (1.6% [one case]). HIV-infected individuals were more likely to have zoonotic genotypes than the HIV-uninfected individuals. Among the C. parvum group, strains clustered distinctly into either human or bovine genotypes regardless of the geographical origin, age, or HIV status of the patients. The intragenotypic variation observed in the C. parvum human genotype was extensive compared to that within the C. parvum bovine genotype group. The variation within genotypes was conserved in all geographical regions regardless of the patients' HIV status. The extensive diversity within genotypes at the 18S rRNA gene locus may limit its application to phylogenetic analyses.     

High-resolution DNA melting curve analysis to establish HLA genotypic identity

High-resolution melting curve analysis is a closed-tube fluorescent technique that can be used for genotyping and heteroduplex detection after polymerase chain reaction. We applied this technique at the HLA-A locus and suggest that this method can be used as a rapid, inexpensive screen between siblings prior to living-related transplantation. At any locus, there are seven general cases of shared alleles among two individuals, ranging from identical homozygous genotypes (all alleles shared) to two heterozygous genotypes that share no alleles. We studied each case using previously typed cell lines to show that identity or non-identity can be determined in all cases by high-resolution melting curve analysis. HLA genotype identity is suggested when two individuals have the same melting curves. Identity is confirmed by comparing the melting curve of a 1:1 mixture with the individual melting curves. Non-identity at the amplified locus changes the heteroduplexes formed in the mixture compared with the original samples and alters the shape of the melting curve. The technique was tested on DNA from a 17-member CEPH family. High-resolution melting curve analysis revealed six different genotypes in the family. The genotype clustering was confirmed by sequence-based typing. Although this technique does not sequence or determine specific HLA alleles, it does rapidly establish identity at highly polymorphic HLA loci. The technique may also prove useful for confirmation of HLA genotypic identity between unrelated individuals prior to allogeneic hematopoietic stem-cell transplantation.