山东淄博地区汉族人群HLA-A、B、DRB1基因频率调查

目的 调查山东淄博地区汉族人群HLA基因多态性的分布。方法 采用序列特异性引物PCR（PCR-SSP）方法,对HLA-A、B、DR位点进行低分辨基因分型;用直接计数法进行抗原频率和基因频率计算。结果 得到一组抗原频率和基因频率资料。结论 山东淄博地区汉族人群HLA基因具有多态性,PCR-SSP方法进行HLA分型的结果准确可靠。     

洛阳地区汉族人群Duffy血型基因型分布

目的调查洛阳地区汉族人群Duffy血型的分布情况。方法收集洛阳本地汉族献血者样本571份,应用血清学方法进行Duffy血型表型鉴定,使用PCR-SSP技术进行基因分型。结果 571份样本中,Duffy血型各种表型分布为:Fy(a+b-)表型523例(91.59%),Fy(a+b+)表型42例(7.36%),Fy(a-b+)表型6例(1.05%),未发现Fy(a-b-)表型。基因频率为:Fya(0.952 7),Fyb(0.047 3)。结论洛阳地区汉族人群Duffy血型分布情况与国内其他地区汉族人群相似,与白种人和黑种人有明显的差别。     

311份脐血样本HLA基因型无法判读原因探讨

本研究探讨影响脐血样本HLA分型结果的因素。采用序列特异性引物PCR反应（sequence—specific priming,SSP）对3II份经过PCR—SS0和PCR—SSP方法仍无法判读HLA型别的脐血进行HLA低分辨重新分型,并对其中7份仍然无法确定HLA型别的脐血样本进行直接测序（sequence—based typing,SBT）,分析无法判读的原因。结果显示：311份脐血样本中99．4％样本不能判定是因为当时HLA分型方法本身的限制性所造成的,0．6％样本不能判定的原因怀疑是脐血中存在母血污染所致。结论：HLA基因分型方法中序列特异性寡核苷酸探针杂交法（sequence—specifi coligonucleotide probe hybridization,SSO）探针设计不足及PCR—SSP等位基因特异性引物的缺乏是影响HLA分型结果无法判读的主要因素。     

RhDel表型与RHEC表型的相关性研究

目的分析广东汉族人群RhD阴性个体中,RhDel表型个体RHEC表型的分布情况和两者的相关性,以及PCR—SSP技术在RhDel基因检测中的应用价值。方法对400份广东汉族人群中血型血清学检测为RhD阴性的标本。采用血型血清学方法对RhDel表型和RHEC表型进行检测,并采用PCR—SSP方法进行RHD1227G〉A基因检测。结果在400份RhD阴性标本中,89例（22．25％）检测到RhDel表型,主要分布在CCee、CeEe和Ccee表型的个体,在CCEe、ccEE、ccEe和ccee表型中未发现。在RhDel表型的标本中,同时检测到RHD1227G〉A基因。结论RhDel表型与RHEC表型具有一定的相关性;应用PCR—SSP技术能够对RhDel基因进行检测;与传统的间接抗人球蛋白实验和吸收放散实验相比,PCR．SSP技术能够快速、便捷筛查RhDel献血者和受血者,避免抗．D抗体引发的同种免疫反应和RhD阴性血液的浪费。     

HLA-DRB1等位基因与中国北方汉族多发性肌炎/皮肌炎的相关性

目的　探讨HLA DRB1等位基因与中国北方汉族多发性肌炎 (PM) 皮肌炎 (DM)的相关性。方法　采用聚合酶链反应 序列特异性引物 (PCR SSP)技术 ,检测中国北方汉族PM DM患者的HLA DRB1等位基因。结果　与 16 8例正常对照组比较 ,在 5 2例PM DM患者中HLA DRB1 0 40x和DRB1 12 0x等位基因频率明显增高 ,且差异有非常显著性 ( χ2 =2 9.80 ,RR =6 .6 1,Pc <0 .0 1;χ2 =2 2 .2 2 ,RR =5 .82 ,Pc<0 .0 1) ;DRB1 0 70x的基因频率也明显高于正常对照组的基因频率 ,且差异有显著性 ( χ2 =10 .6 8,RR =4.48,Pc<0 .0 5 )。 38例DM患者中HLA DRB1 0 40x和DRB1 12 0x等位基因频率明显增高 ,差异有非常显著性 ( χ2 =2 6 .33,Pc<0 .0 1;χ2 =2 0 .82 ,Pc<0 .0 1) ;DRB1 0 70x的基因频率也明显高于正常对照组的基因频率 ,且差异有显著性 ( χ2 =9.6 2 ,Pc<0 .0 5 )。 14例PM患者HLA DRB1 0 40x基因频率也明显增高 ,但经P值校正后无统计学意义 ( χ2 =6 .12 ,Pc>0 .0 5 )。 10例伴间质性肺炎型患者HLA DRB1 12 0x等位基因频率较正常对照组的基因频率明显增高 ,且差异有非常显著性 ( χ2 =12 .5 6 ,Pc<0 .0 1)。结论　PM DM与HLA DRB1 0 40x、 0 70x和 12 0x有显著性相关 ,为揭示PM DM的发病机制中免疫遗传学机制所起的     

HLA-DRB1等位基因与吉林地区汉族白癜风和寻常型银屑病相关性研究

目的：探讨HLA-DRB1等位基因与吉林地区汉族白癜风和银屑病的相关性。方法：采用聚合酶链反应-序列特异性引物（PCR-SSP）分型技术,对82例汉族白癜风患者和80例寻常型银屑病患者HLA-DRB1等位基因进行检测,与86名汉族健康人群进行对照。结果：①白癜风患者组HLA-DRB1＊07和HLA-DRB1＊12等位基因频率比对照组明显增高（P〈0.05）;②银屑病患者组HLA-DRB1＊07等位基因频率高于对照组,HLA-DRB1＊01等位基因频率低于对照组（P〈0.05）。结论：①HLA-DRB1＊07和HLA-DRB1＊12等位基因可能是吉林地区汉族白癜风的易感基因或与易感基因相连锁;②HLA-DRB1＊07等位基因可能是寻常型银屑病的易感基因,HLA-DRB1＊01可能为寻常型银屑病的保护性基因。     

A simplified PCR-SSP method for HLA-A2 subtype in a population of Wuhan, China

HLA-A2 is the most frequent HLA-A allele in all ethnic populations, and an important restriction element for peptide presentation to T cells in infectious disease and cancer. However, the HLA-A2 supertype consisting of up to 75 subtypes, mutation studies and analyses using cytotoxic T lymphocytes suggest the functional relevance of subtype-specific differences in HLA-A2 molecules for peptide binding and T-cell recognition. Therefore, it is necessary for T-cell response study to discriminate the HLA-A2 subtypes and to understand the profile of HLA-A2 allellc distribution in a given population. In this study, we developed a simple, robust approach based on the nested polymerase chain reaction using sequence-specific primers （PCR-SSP） to discriminate 17 HLA-A2 subtypes which cover the most HLA-A2 alleles （〉 99% allele frequency） reported in Chinese, using 15 combinations of 19 allelic specific primers. In the first round of PCR, 3 combinations of 5 primers were used to determine whether the tested sample was HLA-A2 positive, meanwhile the subtypes of HLA-A＊0209 and HLA-A＊0215N were determined for the variant position of these two subtypes is in exon 4 instead of exon 2, 3. Samples of HLA-A2 positive were subtyped in the second round of PCR, using PCR products of the first round as templates. This strategy was applied to test the samples of 78 random HLA-A2 positive individuals for their HLA-A2 subtypes. Those samples were screened for HLA-A2 positive by the first round PCR-SSP from 154 healthy blood donors in Wuhan, China. The subtyping results were verified by using flow cytometric analysis （FCM） with HLA-A2 specific monoclonal antibody BB7.2 and DNA sequencing. The typing results of the samples show 50.7% random individuals in the population carry HLA-A2, HLA-A＊0201 ranks the first （allele frequency = 15.5%）, followed by A＊0207 （5.8%）, A＊0206 （4.7%）, A＊0203 （2.6%）, A＊0210 （0.7%）, and these 5 alleles account for 99.0% HLA-A2 subtypes of allele frequency. Our study indicates that the developed typing method is simple and reliable for HLA-A2 subtyping in Chinese, and the profile of allelic distribution of HLA-A2 subtypes is revealed in the population of Wuhan, China.     

北方汉族人MBL EXON I 52位基因频率调查

目的 初步调查北方地区汉族人MBL52位密码子基因频率的情况。方法 采用MBL 52位密码子基因突变的序列特异性引物聚合酶链式反应即PCR—SSP检测方法。结果 270例健康汉族人52位密码子的野生型为261例，占96．67％；突变杂合型为8例，占2．96％。突变纯合型为1例，占0．37％。结论 基因频率分别为CGT为O．9815；TGT为0．0185。     

Sequence of a new DR12 allele with two silent mutations that affect PCR-SSP typing

A new HLA-DR12 allele has been identified in a European Caucasoid bone marrow donor. The DRB1*12012 allele differs from DRB1*12011 by two silent substitutions at codons 72 and 78, two polymorphic positions used for DNA subtyping of the DR12 serotype. The co-occurence of the two nucleotide changes is unique to the DR12 group and results in a new PCR-SSP typing pattern. The complete HLA type of the donor is A24, A68; B55, B61; Cw*01, Cw*0304; DRB1*12012, DRB1*1402; DRB3*0101, DRB3*0202; DQB1*0301. HLA-DRB1*12012 is a rare allele as it occurs in < 0.2% of DR12 donors.     

Distribution of the HLA-A9 antigen family (A23, A24 and A2403)

Characterization of 50 local anti-HLA-A9 (A23 and/or A24) sera against the HLA-A2403 antigen allowed the retrospective assignment of A2403 and the verification of A23 and A24 specificities in a panel of 9196 volunteer bone marrow donors. The six A2403 positive subjects identified were confirmed by PCR using four sequence-specific primer mixtures. Population analysis showed the distribution and HLA-A/B linkage disequilibrium of HLA-A23 and A24 to be typical of a Northern European Caucasoid population. HLA-A2403 has a phenotype frequency of 0.00065 (gene frequency - 0.00033) and is in linkage disequilibrium with HLA-B63 and possibly B35.