HLA-DRB1基因多态性与HBV宫内感染的相关性

目的 探讨HLA-DRBl等位基因多态性与HBV宫内感染的关系.方法 选择HBsAg阳性孕妇分娩的新生儿中发生HBV宫内感染的母亲21例(A组),而未发生宫内感染的母亲46例(B组)以及正常母亲42例(C组),采用聚合酶链反应-序列特异性引物(PCR-SSP)技术扩增HLA-DRB1四对等位基因,计算各刑别出现频率.结果 (1)HLA-DRB1*11在A组的等位基因频率明显高于B组(38.1% vs 13.0%)(P<0.05);(2)HLA-DRB1*13、HLA-DRB1*03在A、B两组的等位基因频率明显低于C组(4.5% vs 33.3%,9.0%vs 28.6 0A)(P<0.01):(3)HLA-DRB1*07等位基因频率在A、B、C三组间差异无显著性.结论 HLA-DRB1*11与HBV宫内感染有关;HLA-DRB1*03、13可能与HBV清除有关.     

河南地区汉族人群药物代谢酶CYP2C19，NAT2和TPMT基因多态性分析(英文)

目的：了解细胞色素P450(cytochromes P450,CYP)2C19,N-乙酰基转移酶2(arylamine N- acetyltransferase 2,NAT2)和硫嘌呤甲基转移酶(thiopurine S-methyltransferase,TPMT)基因常见的遗传多态性在河南地区汉族人群中的分布及其频率。方法：应用聚合酶链反应-限制性片段长度多态性分析(PCR-RFLP)对210名河南地区汉族人群的CYP2C19突变基因(*2和*3)、NAT2突变基因(*6和*7)和TPMT突变基因(*3A,*3B和*3C)进行检测。用聚合酶链反应-等位基因特异性扩增(PCR-ASA)对NAT2突变基因(*5)和TPMT突变基因(*2)进行检测。结果：CYP2C19*2和*3等位基因分布频率分别为34．76%和6．4%,同时携带2个等位突变基因的慢基因型频率占14．8%。NAT2*4(wt),*5(341C),*6(590A)和*7(857A)等位基因分布频率分别为59．1%,4．1%,26．4%和9．5%,慢基因型分布频率占19．5%。TPMT*3C等位基因分布频率为1．2%,未发现TPMT*2,TPMT*3A或TPMT*3B。结论：CYP2C19,NAT2和TPMT基因常见的遗传多态性在汉族人群中的分布及其频率与白人存在明显差异,这将有助于我国汉族人群临床药动学研究和给药剂量的确定。     

癫痫患者CYP2D6~* 10基因多态性分析

目的:通过比较本地区健康人与癫痫患者CYP2D6*10基因多态性的差异,探索CYP2D6*10基因多态性与癫痫易感的关系。方法:应用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)方法检测健康人和癫痫患者CYP2D6*10的基因型分布及等位基因频率,通过χ2检验来比较健康人和癫痫患者的基因型分布差异和等位基因频率差异。结果:健康人和癫痫患者的基因型分布及等位基因频率差异有显著性和高度显著性(P<0.05和P<0.01)。结论:癫痫病的发生与CYP2D6*10的碱基变异具有相关性。     

河北汉族HLA-A基因多态性及其分布特征

目的 了解河北省汉族人群人类白细胞抗原HLA-A等位基因多态性及分布特征.方法 采用序列特异性寡核苷酸探针聚合酶链反应(PCR-Sequence Specific Oligonucleotide Probing,PCR-SSOP)方法,对5 908名河北汉族捐献造血干细胞健康志愿者的HLA-A等位基因作低分辨基因分型检测,采用方根法计算HLA-A基因频率,并进行群体比较.结果 河北汉族人群中共检出18个HLA-A基因,呈现较高的基因多态性,包括过去很少被检测到的HLA-A25,A74,其中A座位最常见基因依次为:A* 02、A*11、A*24、A*01、A*30、A*03、A*33.结论 河北汉族人群HLA-A基因分布有自己的特点,各等位基因频率介于南北汉族之间,但更接近于中国北方汉族人群,符合地域分布特征.     

HLA-DRB1、HLA-DQB1基因多态性与云南汉族人群耐多药肺结核病的关联研究

摘要：目的:探讨云南地区汉族人群HLA-DRB1、HLA-DQB1基因多态性与耐多药肺结核病发生的关系。方法:收集2017～2018年在我院就诊的云南省汉族耐多药肺结核（MDR-PTB）患者300例作为观察组;门诊体检健康人群300例作为对照组。应用聚合酶链式反应-直接测序基因分型法（PCR-SBT）,检测两组人群HLA-DRB1、HLA-DQB1等位基因分布频率。结果:共检测到25个HLA-DRB1等位基因,17个HLA-DQB1等位基因。其中观察组的HLA-DRB1*07:01等位基因频率显著性高于对照组[18.7%vs.13.5%,P=0.007,OR=1.471,95%CI（1.077,2.008）],观察组的HLA-DQB1*03:01等位基因频率显著性高于对照组[27.8%vs.20.3%,P=0.002,OR=1.511,95%CI（1.157,1.974）],其余等位基因两组间无明显差异（P>0.05）。结论:HLA-DRB1*07:01和HLA-DQB1*03:01等位基因可能是云南汉族人群耐多药肺结核的易感基因。     

广西壮族自治区妊娠期糖尿病与人类白细胞抗原Ⅱ类基因多态性的关系

目的探讨广西壮族自治区人类白细胞抗原(HLA)-Ⅱ类基因多态性与妊娠期糖尿病的相关性。方法选取妊娠期糖尿病(GDM)孕妇100例作为试验组,100例糖耐量正常的孕妇作为对照组,采用序列特异性引物聚合酶链反应(PCR)对2组孕妇的HLA-Ⅱ类基因进行检测和分析。结果 2组孕妇经基因检测后,GDM组等位基因DQA1*0301、DRB1*0301和DRB1*1302的频率显著高于对照组,差异具有统计学意义(P<0.05);DQA1*0103、DQA1*0501和DQB1*0601等位基因的频率低于对照组,但差异无统计学意义(P>0.05),DQB1*0201、DQB1*0302、DQB1*0602等位基因的频率高于对照组,但差异也无统计学意义(P>0.05)。结论在广西壮族自治地区,HLA-Ⅱ类等位基因与GDM的发病存在相关的联系,等位基因DRB1*0301、DRB1*1302和DQA1*0301是妊娠期糖尿病的易感基因。     

中国东北地区汉族癫痫患者HLA-B*1502等位基因分布频率

目的研究中国东北地区汉族癫痫患者HLA-B*1502等位基因分布的频率。方法用聚合酶链反应DNA序列分析法,对125例中国东北地区汉族癫痫患者DNA标本进行HLA-B基因分型。结果 125例DNA标本中,共检测出34种不同的HLA-B等位基因。其中,2例患者携带HLA-B*1502等位基因。HLA-B*1502等位基因在中国东北地区癫痫患者中的分布频率为1.6%。其他分布频率较高的等位基因分别为HLA-B*4001、B*4601、B*4801、B*1302和B*4002。结论东北地区癫痫患者中HLA-B*1502等位基因分布频率(1.6%)与其在北京地区健康汉族人群中的分布频率近似(1.5%);但远低于其在中国中部、西部及南部地区的分布频率。     

乙型肝炎表面抗原阳性母亲新生儿乙型肝炎病毒宫内传播影响因素研究

目的探讨乙型肝炎表面抗原(HBsAg)阳性母亲新生儿发生乙型肝炎病毒(HBV)宫内传播的影响因素。方法收集2011年7月至2013年7月在太原市第三人民医院妇产科分娩的HBsAg阳性孕妇及其新生儿396例。以是否发生HBV宫内传播将新生儿分为宫内传播组和非宫内传播组,分析母亲年龄、孕期HBV血清标志物水平、分娩方式、新生儿性别与新生儿HBV宫内传播的关系。统计分析采用χ2检验、趋势χ2检验和非条件Logistic回归模型。结果单因素分析显示,母亲孕期乙型肝炎e抗原(HBeAg)阳性、HBV DNA阳性、HBeAg/HBV DNA双阳性和经剖宫产分娩与新生儿HBV宫内传播的发生有关(P<0.05);母亲年龄、新生儿性别与宫内传播的发生无关(P>0.05)。多因素分析显示,HBeAg/HBV DNA双阳性(OR=3.662,95%CI为1.929⁶.951)、经剖宫产分娩(OR=0.184,95%CI为0.0906.951)、经剖宫产分娩(OR=0.184,95%CI为0.090⁰.375)被引入回归方程。趋势χ2分析后显示,HBV DNA≥1×107copies/ml时新生儿HBV宫内传播发生率明显上升。结论经剖宫产分娩是HBsAg阳性母亲新生儿HBV宫内传播的保护因素,而母亲孕期血清HBeAg/HBV DNA双阳性和HBV DNA≥1×107 copies/ml时,新生儿HBV宫内传播发生的可能性较大。     

乳腺癌组织p21WAF1基因多态性及其与预后的关系

目的探讨乳腺癌组织p21^WAF1 DNA多态性及其与预后的关系。方法采用聚合酶链反应单链构象多态性技术(PCR-SSCP)和免疫组化S-P法检测100例乳腺癌组织p21^WAF1 DNA多态性和蛋白表达，并根据随访结果行预后分析。结果100％(18／18例)的具有p21^WAF1 DNA多态性的乳腺癌组织p21^WAF1蛋白表达阳性，39％(32/82)的无p21^WAF1 DNA多态性乳腺癌组织蛋白表达阳性，p21^WAF1 DNA多态性组明显高于无多态性组(X^2=21．95，P＜0．01)；p21^WAF1 DNA多态性与p21^WAF1蛋白表达呈正相关(rs=0．576，P＜0．01)。单因素预后分析显示p21^WAF1 DNA多态性组生存期低于无多态性组(X^2=6．02，P＜0．01)。结论乳腺癌p21^WAF1 DNA多态性可能是造成乳腺癌患者生存期缩短的一个重要因素。     

中国汉族和维吾尔族药物代谢酶CYP3A4、CYP2C9、CYP2C19、CYP2D6基因多态性分析

目的对中国汉族、维吾尔族健康人群CYP3A4、CYP2C9、CYP2C19、CYP2D6进行基因多态性分析,并对汉族和维吾尔族人群等位基因频率和基因型频率进行比较。方法聚合酶链反应一限制性片段长度多态性(PCR-RFLP)法对CYP3A4、CYP2C9、CYP2C19、CYP2D6进行分型。结果汉族、维吾尔族健康人群CYP3A4*5等位基因频率为0,CYP3A4*18等位基因频率分别为0.183 8、0.140 2;CYP2C9*2等位基因频率分别为0.011 0、0.095 8,CYP2C9*13等位基因频率分别为0、0.002 3;CYP2C19*2等位基因频率分别为0.386 0、0.324 8,CYP2C19*3等位基因频率分别为0.051 5、0.021 0;CYP2D6*10等位基因频率分别为0.573 5、0.224 3。结论本研究在汉族、维吾尔族健康人群中未发现CYP3A4*5等位基因。汉族、维吾尔族健康人群CYP3A4*18、CYP2C9*13、CYP2C19*2、CYP2C19*3等位基因频率差异均无统计学意义。维吾尔族健康人群CYP2C9*2等位基因频率远高于汉族(P<0.01);CYP2D6*10等位基因频率远低于汉族(P<0.01),存在民族差异。     

乙型肝炎病毒表面抗原阳性孕妇及其新生儿外周血中乙型肝炎病毒的研究

目的了解乙型肝炎病毒表面抗原(HBsAg)阳性孕妇及其新生儿外周血乙型肝炎病毒(HBV)的感染状况。方法酶联免疫吸附试验(ELISA)法检测新生儿外周血HBsAg;巢式聚合酶链反应(nPCR)检测孕妇及其新生儿外周血HBV-DNA;选择性聚合酶链反应(sPCR)检测孕妇及其新生儿外周血单个核细胞(PBMC)中HBV-DNA,三项指标任一项阳性即判定为新生儿HBV宫内感染。结果HBsAg阳性孕妇血清HBV-DNA阳性率为43.1%(53/123),PBMCHBV-DNA阳性率为30.8%(37/123);新生儿血清HBsAg阳性率为6.5%(8/123),HBV-DNA阳性率为19.5%(24/123),PBMCHBV-DNA阳性率为26.0%(32/123)。新生儿任一项阳性者50例,合计宫内感染率40.6%(50/123)。结论检测PBMCHBV-DNA对血清中HBV-DNA检测进行补充,可以比较准确地反映HBsAg阳性孕妇新生儿宫内HBV感染的状况。     

CYP2D6 genetic polymorphism in South Indian populations

This study aimed to determine the prevalence of genetic polymorphism in the CYP2D6 gene, which codes for the polymorphically expressed CYP2D6 drug-metabolizing enzyme. The common variants CYP2D6 *2, *3, *4, *5, *10, *14, and *17 were studied in the populations (n=447) of the four South Indian states namely Tamilnadu (TN), Kerala (Ker), Karnataka (Kar) and Andhra Pradesh (AP). Genetic polymorphisms were identified using polymerase chain reaction (PCR) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) based methods. Differences in frequencies of CYP2D6 polymorphism between each South Indian state were statistically compared, and also the frequency of South Indian population as a whole in relation to other major populations. The CYP2D6*2 allele was the most frequent variant (34.8%), followed by the *10 allele (10.2%). The *4 and *5 alleles occurred at 7.3% and 1.9% respectively. The *3, *14 and *17 alleles were not detected in the study. The *1/*2, *1/*1 and *2/*2 genotypes were the most common CYP2D6 genotypes, representing 32.7%, 19.4% and 11.8% of the South Indian population. Genotypes that predict poor metabolizer phenotype i.e. *4/*4 and *4/*5 were found at 0.6% in South Indian population. The genetic composition at the CYP2D6 locus in South Indians is distinct from Caucasian, African and even other Asian (Chinese and Japanese) populations.     

Prevalence of DRB1*01 and DRB1*04 alleles in a group of patients with rheumatoid arthritis living in Liguria

A major component of genetic susceptibility to rheumatoid arthritis (RA) appears to be explained by inheritance of HLA-DRB1 alleles. Multiple HLA-DRB1 alleles (DRB1*0401, *0404, *0405, *0408, *0101, *102, *1001 and *1402) encoding a shared epitope at amino acid positions 70-74 are associated with susceptibility to RA. There is ethnic variation in the clinical expression of RA and in both the frequency and type of HLA-DRB1 alleles carrying the shared epitope. We evaluated the prevalence of the alleles of HLA-DRB1 locus encoding for SE in 42 outpatients with RA attending the Rheumatology Center of the University of Genoa, Bruzzone Institute, and living in Liguria. A control group was composed of Italian marrow donors. DNA genotyping was performed using a low-resolution polymerase chain reaction technique for characterization of the families of HLA-DRB1 alleles for each of the 42 patients studied. Subsequently, subjects with *01 and *04 haplotype were tested with high-resolution HLA-DRB 1 typing to characterize the *01 and *04 alleles. No statistically significant differences were found in the prevalence of RA-associated single alleles *01 and *04 in the study group or in the control group. In contrast, the sum of susceptibility *04 alleles studied by resolution typing was strongly related to RA in the study group in comparison with the control group.     

Optimization of cytochrome P4502D6 (CYP2D6) phenotype assignment using a genotyping algorithm based on allele frequency data

Cytochrome P4502D6 (CYP2D6) is a highly polymorphic gene locus with > 50 variant alleles which lead to a wide range in enzymatic activity. So called poor metabolizers are carriers of any two non-functional alleles of the CYP2D6 gene. CYP2D6 genotyping is cumbersome and the question of how much genotyping is necessary for an accurate phenotype prediction is still debated. The goal of this study was to determine the optimum amount of genotyping required to accurately predict the phenotype at a reasonable cost in a white North American population. To address this issue, we designed a polymerase chain reaction (PCR)/restriction fragment length polymorphism-based genotyping strategy to detect 'key' mutations linked to extensive metabolizer or poor metabolizer associated alleles in combination with extra-long PCR (XL-PCR). All mutations with the exception of gene deletions and duplications are detectable by simple restriction digestion analysis and agarose gel electrophoresis. In addition, we utilized a genotyping algorithm based on our own and published allele frequency data and phenotype analysis to calculate the probability of a correct genotype (and thus, phenotype) assignment. As little as one XL-PCR reaction followed by a maximum of six reamplification reactions allows an accurate prediction of an individual's genotype to 99.15%. As few as four reamplification reactions identify 97.9% of poor metabolizer individuals. We evaluated our model in 208 white North Americans by testing for the presence of 'key' mutations linked to CYP2D6*2, *3, *4, *6, *7, *8, *9, *10, *11, *12, *15, *17 and *18 alleles and the *5, *13 and *16 gene deletions. For all individuals, the correct phenotype has been predicted. Discordant phenotype assignment occurred in only two individuals which subsequently was attributed to CYP2D6 inhibition by concomitant drug therapy.     

The <Emphasis Type="Italic">CYP2D6</Emphasis> gene locus in South African Coloureds: unique allele distributions,novel alleles and gene arrangements

Objective The highly polymorphic CYP2D6 gene locus has been extensively scrutinised in the major ethnic populations, but little is known about the locus for many indigenous and unique admixed populations, including the Coloureds of South Africa. This study aimed at characterising the CYP2D6 gene locus in Coloured subjects and predict their phenotype status. Methods CYP2D6 genotyping was performed on 99 Coloured subjects by long-range polymerase chain reaction (PCR) and PCR restriction fragment length polymorphism (RFLP). Testing included 25 allelic variants as well as gene duplications. Novel alleles were cloned and sequenced. A novel strategy for CYP2D7/2D6 hybrid gene detection is described. Results Thirteen alleles had a frequency of = 1%, three were infrequent (<1%) and 16 of the tested alleles were not detected. CYP2D6*5 had a frequency of 17.2%, one of the highest ever observed in any population. Two novel alleles, CYP2D6*64 and *65, were identified. In addition, four samples carried CYP2D7/2D6 hybrid genes, of which one matched the CYP2D6*66 allele of a resequenced Caucasian control subject. CYP2D6*66 is similar to CYP2D6*16, but its putative recombination point is further upstream. Genotyping identified three poor metabolisers (3%; predicted incidence 6.6%), and 12% of the population had an activity score of 0.5 indicative of intermediate metabolism. Conclusions The Coloured population has a unique composition of alleles and a distinct frequency distribution. The preliminary data presented suggest decreased CYP2D6 activity compared with Caucasians. Follow-up studies including both genotyping and phenotyping will need to be conducted to further assess the relationship between genotype and phenotype in this population of complex ancestry.     

Genetic polymorphism of cytochrome P450 2C9 in a Caucasian and a black African population

AIMS: CYP2C9 is a major enzyme in human drug metabolism and the polymorphism observed in the corresponding gene may affect the therapeutic outcome during treatment with several drugs. The distribution of variant CYP2C9 alleles was therefore investigated in an Italian and an Ethiopian population. METHODS: Allele-specific PCR analysis was carried out in order to determine the frequencies of the two most common variant alleles, CYP2C9*2 and CYP2C9*3 in genomic DNA isolated from 157 Italians and 150 Ethiopians. RESULTS: The frequencies of CYP2C9*1 (80%), CYP2C9*2 (11%) and CYP2C9*3 (9%) found in the Italian population were similar to other Caucasian groups. However in the Ethiopian population CYP2C9*1, CYP2C9*2 and CYP2C9*3 were present at a frequency of 94, 4 and 2% respectively. The 95% confidence intervals in CYP2C9*1, CYP2C9*2 and CYP2C9*3 between Italians and Ethiopians were 0.098, 0.176, 0.040, 0.098 and 0.040, 0.098, respectively. CONCLUSIONS: Our results indicate that the Ethiopian population has a unique relative distribution of the CYP2C9 alleles, which is not similar to any other ethnic group hitherto described.     

The impact of CYP2C9 genetics and oral contraceptives on cytochrome P450 2C9 phenotype.

CYP2C9-dependent drug metabolism is subject to large interindividual variation. To some extent, this is explained by genetic polymorphism with expression of enzyme variants that differ in catalytic activity. The aim of this study was to characterize the variation in CYP2C9 phenotype in relation to genotype, with further analysis of the CYP2C9 gene in metabolic outliers. A study population of 126 healthy white subjects were recruited and genotyped for the variant alleles, CYP2C9*1-3. In CYP2C9 phenotyping with losartan, three subpopulations were distinguished that differed in the number of CYP2C9*3 alleles (0, 1, or 2). A three-fold higher metabolic ratio (MR; urinary losartan/carboxymetabolite) was found comparing CYP2C9*1/*3 (n = 20) to CYP2C9*1/*1 (n = 81), but there was considerable variation within each genotype. Subjects genotyped as CYP2C9*1/*1, but with an unexpectedly slow oxidation of losartan, were selected for DNA-sequencing analysis of the CYP2C9 gene. Interestingly, single nucleotide polymorphisms (SNPs) could not be identified either in the 5'-flanking region, the nine exons, or exon-intron boundaries. However, sequencing of the CYP2C9 gene was also carried out in patients genotyped as CYP2C9*1/*1 but with an exceptionally low steady-state clearance of S-warfarin. Here, five different SNPs were identified. In further analysis of the healthy volunteers, it became evident that women on oral contraceptives (OCs) had slower oxidation of losartan (MR of losartan: 1.7) than women without OCs (MR of losartan: 0.86). This novel finding was not explained by a different frequency of variant alleles. In summary, CYP2C9 genotype and oral contraceptives both contribute to a large interindividual variation in CYP2C9 activity.     

短串联重复序列VWA在太原汉族人群的法医应用研究

目的　获得短串联重复序列 VWA(nt16 40 - 1794)在太原汉族人群的群体遗传学数据 ,探讨其在法医学中的应用。方法　用 TKM法对 2 14名太原地区无关汉族人群抽提血液 DNA。扩增特异性片段 ,含 7m ol/ L 尿素的聚丙烯酰胺凝胶垂直电泳 ,银染显色分析其多态性。结果　该位点频率分布符合 H- W平衡定律。共检出 8个等位基因 ,长度在 134 - 16 2 bp之间 ,频率分布 ,VWA* 13- 0 .0 0 7;VWA* 14- 0 .194;VWA* 15 - 0 .0 2 8;VWA* 16 -0 .173;VWA * 17- 0 .2 78;VWA * 18- 0 .140 ;VWA * 19- 0 .10 3;VWA * 2 0 - 0 .0 14。个体识别率 :0 .932 ,杂合度0 .82 1,多态信息含量 0 .814,非父排除率 0 .6 0 3。本文调查的频率同大部分欧洲人及美国黑人分布差异有显著性。同其他中国人差异无显著性。结论　 VWA基因座在太原汉族人群中有较高的杂合度、个体识别率及非父排除率 ,是高识别能力遗传标记 ,在法医学和群体遗传学有很高的应用价值。     

CYP2D6 PCR基因型与DXT表型和基因芯片检测的比较

目的：为了评价CYP2D6的基因型和表型的联系以及基因芯片在CYP2D6多基因分析中的应用。方法：242健康志愿者，口服dextromethorphan后收集认测定其代谢率，收集20ml血提取DNA，并通过基因特异性PCR和／（或）基因芯片分析CYP2D6＊2－＊11，＊17和多拷贝CYP2D6基因，其中5个基因（＊3，＊4，＊6，＊7和＊9）用PCR和CYD450基因芯片同时分析。结果：CYP2D6基因型比表型更富有信息和更能反映CYP2D6酶的表达。CYP2D6＊3，＊4，＊6，＊6和＊9的基因检测在CYP450基因芯片和基因特异性PCR中显示高度的一致性。结论：基因芯片在检测基因多位点的多基因中是一个有发展前途和可靠的方法。     

武汉汉族人群DYS446基因座的遗传多态性

为研究 Y染色体上 DYS44 6基因座多态性 ,应用 PCR扩增 DYS44 6基因座、变性聚丙烯酰胺凝胶电泳、银染的方法检测了 10 7份汉族男性无关个体血样本、10份女性血样本和 2例已确认亲子关系的父子血样本。结果显示 ,10 7例男性血样本有相应的PCR产物 ,共检测到 8个等位基因 ,GD值为 0 .764 9;在 10例女性中 ,无相应的 PCR产物 ;2例已确认父子关系的基因型一致。提示DYS44 6基因座是个人识别和亲子鉴定的较好遗传标记。