石家庄地区初筛Rh(D)阴性献血者D变异体表型分析

目的研究石家庄地区初筛为Rh(D)阴性献血者中D变异体表型的分布。方法263份初筛为Rh(D)阴性的血样用间接抗球蛋白试验筛选并确认弱D/部分D表型,然后用吸收放散试验从间接抗球蛋白试验阴性的血样中筛选并确认Del表型,Rh因子用常规血清学试验检测。结果间接抗球蛋白试验共筛出弱D/部分D表型16份,其中RhCcee3份(18.75%)、RhCcEe1份(6.25%)和RhccEe12份(75.00%);吸收放散试验共筛出Del表型59份,其中RhCCee5份(8.47%)、RhCcee51份(86.44%)和RhCcEe3份(5.08%)。结论石家庄地区初筛为Rh(D)阴性献血者中,6.08%为弱D/部分D表型,22.43%为Del表型,且与RhCE表型有关。     

广东省中山地区临床初检RhD阴性人群中D变异体分布及特征分析

目的　研究分析中山地区初检 RhD阴性人群中 D变异体血清学和基因分型特征。方法　研究共收集 2017年 12月～ 2019年 2月中山市人民医院门诊及住院病人的血液样本 24 286例,采用微柱凝胶卡法对其中 RhD阴性样本进行初筛;再经间接抗人球蛋白试验（ indirect antiglobulin test, IAT）确认弱 D或部分 D变异型;然后用吸收放散试验对剩余样本进行 Del表型筛选。同时,通过聚合酶链反应 -序列特异性引物（ polymerase chain reaction-sequence speci.c primmer, PCR-SSP）技术对初筛 RhD阴性的样本进行 RHD等位基因分型以及血清学方法对初筛阴性样本进行 C, c, E和 e抗原表型的检测。结果　在 24 286例血液样本中初筛共检出 102例阴性样本,中山地区初检阴性比例约为 0.42%。经 IAT确认试验共鉴定出 7例弱 D或部分 D血液样本（ 6.86%）,且基因型表现多样,但未见亚洲地区常见的弱 D15和弱 D12表型。接下来的放散试验检测出 25例 Del型,通过 PCR-SSP基因分型技术又检出 3例漏检的 Del型,Del型在初检阴性样本中占比 27.45%,PCR-SSP结果显示该研究中所有 Del型血液样本等位基因均为 RHD1227A。该研究纳入的 102例初检阴性血液样本的 RhCcEe表型分布符合 Hardy-Weinberg遗传平衡 (χ2=2.625,P>0.05),表示相应等位基因所占比例在遗传中保持不变。在 Del型变异体中,除 Ccee和 CCee表型外,未见其他表型。结论　中山地区人群 RhD变异体有丰富的类型和不同分子机制,其中 Del型占比较高,血清学与基因分型结合可提高 D变异体的鉴定能力,防止漏检情况的发生。     

Rh（D）变异体的研究进展

Rh（D）变异体的检测在临床Rh血型检测中越来越受到重视,本文将总结分析D变异体的研究进展及国内研究现状,为基因技术检测Rh（D）变异体提供理论依据。     

无锡地区献血者D变异体的检定结果

目的准确地检定D变异体,并探讨其在安全输血中的意义。方法通过AS-PCR方法测定RHD1227A等位基因,检定Del血型;通过血清学方法检出D弱反应性标本,然后通过PCR方法鉴定部分D和弱D。结果 351份盐水抗-D阴性标本,检出Del32例,部分D 3例,弱D 4例。结论准确检定D变异体对制定安全有效的临床输血策略具有重要意义。     

华北地区汉族人群Rh(D)抗原弱表现型个体的分子遗传机制研究

目的探讨汉族人群非血缘关系Rh(D)抗原弱阳性个体的血清学表型及分子遗传机制。方法采用常规血清学技术从非血缘关系随机献血者中筛检Rh(D)抗原弱阳性个体(包括弱D型、部分D型),对其进行Rh D、C、c、E、e抗原表型的检测;采用序列特异性引物-聚合酶链反应(PCR-SSP)方法同时检测其RHD基因和RHCE基因;测序分析RHD基因全长编码区序列;同时通过特异性PCR技术测定其RHD合子型。结果血清学试验证实为D抗原弱阳性表型的有32例个体,占无关供者人群比率为0.015%,其中18例个体为弱D15型(845G>A),1例为弱D12型(830G>A),1例为携带DEL等位基因(1227G>A)的弱D型,8例为部分D表型中的DⅥⅢ型(RHD-CE(3-6)-D),1例为部分D表型中的DⅤa(Hus)(RHD-CE(5)-D),3例标本10个外显子检测均未见异常。Rh小因子检测有3种表型CcEe(4例)、Ccee(10例)、ccEe(18例),其血清学与分子生物学检测一致。RHD杂合性试验鉴定显示仅4例标本为纯合型RHD+/RHD+,其余为杂合型RHD+/RHD-。结论汉族人群D抗原弱阳性比率明显少于高加索人,汉族人群D弱表现型中,弱D15型频率最高;部分D的弱表现型中,DⅥⅢ型占主要比例。     

Rh血型D放散型(D~(el))

<正>Rh血型系统是人类红细胞30个血型系统中最为复杂的1个抗原系统,在临床输血中其重要性仅次于ABO血型。Rh血型系统包括50多个抗原,与临床密切相关的抗原主要     

贵阳市无偿献血人群RhD变异体的基因型分析

目的研究分析RhD抗原变异体的血清学表型和基因型。方法收集贵阳市159名无偿献血者RhD盐水法初筛阴性的标本,采用间接抗球蛋白试验(IAT)筛出部分D型和弱D型抗原,用乙醚吸收放散试验检出Del型抗原,并对Rh表型(C、c、E、e抗原)进行血清学检测。同时运用序列特异性引物-聚合酶链反应(PCR-SSP)对RhD变异型D抗原外显子进行扩增和选择性测序以判定其确切型别。结果 159例RhD初筛阴性标本中检出RhD变异型53例(33.3%),包括Del1227GA、3GA 46例(28.9%)、部分D DVI typeШRHD-CE(3-6)-D 3例、弱D15 845 GA 3例、弱D24 1013TC 1例。Rh表型结果显示,ccee 65例、Ccee 79例、CCee 11例、ccEe和CcEe各2例。结论贵阳市无偿献血人群中RhD变异体的基因型呈现多态性,其中主要为Del型。     

RHD* 1227A等位基因在辽宁汉族Rh阴性人群和随机人群中的频率分析

目的 研究调查辽宁地区汉族RHD*1227A等位基因在Rh阴性人群和随机人群中的频率.方法 采用吸收放散实验筛查Del表型,RHD基因特异聚合酶链反应-序列特异性(PCR-SSP)筛查RHD* 1227A等位基因,RHD全编码区的核苷酸序列分析方法确认RHD* 1227A等位基因,杂交合子技术检测RHD基因的杂合性.结...     

聊城市Rh血型表型库的建立

目的了解聊城市献血者Rh不同表型、RhD(-)、部分D、弱D和Del分布,建立Rh血型表型库。方法对聊城市2009年2月～2011年5月的85 381名次初次无偿献血者进行RhD初筛,初筛阴性者再进行RhD(-)确认和表型检测,随机抽取18 317名次RhD(+)无偿献血者进行表型检测,并建立Rh血型系统表型数据库。结果 Rh D(-)211人,弱D 19人,Del82人;RhD(-)献血者各表型中dccee比例最高,其后依次为dCcee和dccEe;RhD(+)献血者各表型中DCCee比例最高,其后依次为DCcEe和DCcee。结论研究无偿献血人群Rh血型分布,建立Rh血型表型数据库,对保证合理计划用血及应急供血具有重要的临床意义。     

Rh血型系统弱D及部分D研究进展

Rh血型系统弱D及部分D具有不同的分子遗传机制,形成弱D的氨基酸替换主要位于胞内和跨膜区域,表现为抗原位点数减少,但抗原表位数目基本不变。形成部分D（partial D）的氨基酸变异主要位于胞外区域,表现为缺失一个或多个D抗原表位,这也是部分D易产生抗体的原因所在。本文主要综述了近年来Rh血型系统弱D及部分D相关研究进展。     

Molecular basis of D variants between Uigur and Han blood donors in Xinjiang

summary .  The D antigen is the most antigenic in the Rh blood group system. Variation in the D antigen may have the potential to cause alloimmunization. D variant may have different molecular backgrounds in people of different ethnic groups. The aim of this study was to investigate serological and molecular differences related to the D antigen among Chinese ethnic groups. Blood samples of six different races in Xinjiang were screened for D variants using serological test. The suspected D variants were further analyzed by using polymerase chain reaction and sequencing to determine the RHD genotype. Fourteen D elute phenotypes (DELs), included 11 Han, 2 Uigur and 1 Hui, were detected together with two weak Ds, one in the Han and one in the Uigur. The 14 DELs possessed the RHD (K409K) allele. The weak D found in the Han was of type 15, but the Uigur phenotype was of weak D type 5. Our results suggest that the Uigur population has both Han and Caucasian characteristics in the Rh blood group system, but the RHD genotypes of other minorities settled in China need to be further studied. A different strategy for Rh typing based on ethnic specificity should be used to detect D variants.     

Frequency of RHD variants in serologically weak D Turkish blood donors

BackgroundRhD typing has remained of primary importance, as being the leading cause of hemolytic disease of the newborn. Among Rh system’s 55 blood group antigens, RhD is the most immunogenic. We aimed with this study to determine weak D/partial D variant frequency in blood donors who were admitted to our blood center and have serologically designated blood group weak D.Materials and methodsWe screened blood donors who admitted between 2011 and 2017 to our blood center. Sixty-seven serologically weak D phenotyped donors have participated in the study. These donors' samples were studied further by Polymerase Chain Reaction Sequence- Specific Primers (PCR-SSP) for determining D variants.ResultsWeak D phenotype was detected in 228(0.12 %) out of 177,554 donors. Sixty-seven of them agreed to take part in the study. The frequency of weak D and partial D was 68.7 % (n = 46), and 22.4 % (n = 15), in order. The most encountered weak D and partial D variant was type 15 and DFR type, respectively.ConclusionsThe prevalence of serologically weak D phenotypes varies by race and ethnicity. Turkey is a country covering a mixture of European and Asian DNA with different ethnic groups. Thus, our research as giving the overall distribution of RHD variants from the largest city of Turkey, which may reflect the general ethnic background of the country, would help to the establishment of a databank for blood banking. This paper is the first molecular study on RHD variants in Turkey. New molecular research would be more reliable and precise.     

Systematic RHD genotyping in Brazilians reveals a high frequency of partial D in transfused patients serologically typed as weak D

BackgroundThe discrimination between weak D types and partial D can be of clinical importance because carriers of partial D antigen may develop anti-D when transfused with D-positive red blood cell units. The aim of this study was to determine by molecular analysis the type of D variants among Brazilian patients requiring transfusions with serologic weak D phenotypes.Material and methodsSamples from 87 patients (53 with sickle cell disease, 10 with thalassemia and 24 with myelodysplastic syndrome), serologic typed as weak D by manual tube indirect antiglobulin test or gel test were first RHD genotyped by using the RHD BeadChip Kit (BioArray, Immucor). Sanger sequencing was performed when necessary.ResultsRHD molecular analysis revealed 32 (36.8 %) variant RHD alleles encoding weak D phenotypes and 55 (63.2 %) alleles encoding partial D antigens. RHD variant alleles were present in the homozygous state or as a single RHD allele, one variant RHD allele associated with the RHDΨ allele, or two different variant RHD alleles in compound heterozygosity with each other in 70 patients, 4 patients and 13 patients, respectively. Alloanti-D was found in 9 (16.4 %) cases with RHD alleles predicting a partial D.DiscussionThe frequency of partial D was higher than weak D types in Brazilian patients serologically typed as weak D, showing the importance to differentiate weak D types and partial D in transfused patients to establish a transfusion policy recommendation.     

D抗原阳性个体及无效等位基因携带者RHD基因数目测定

目的　测定Rh血型D抗原阳性个体的RHD基因数目。方法　采用聚合酶链式反应 限制性片段长度多态性 (PCR RFLP)技术 ,扩增 10名D阳性、5名弱D、2 5名Del表型和 12名携带无效等位基因的Rh阴性个体的RH基因座位下游盒子和融合盒子序列 ,产物经DNA限制性内切酶PstI消化后电泳 ,以 5份经血清学和序列分析基因分型方法确认的D阴性样本作为RHD-/RHD-纯合子对照 ,鉴定RHD+ /RHD+ 或RHD+ /RHD-基因型。结果　 10名D阳性个体均为RHD+ /RHD+ 纯合子、5份弱D样本及 12名携带无效等位基因的Rh阴性个体全部为RHD+ /RHD-杂合型。 2 5名Del表型个体中 9人为RHD+ /RHD+ 纯合型 (36 % ) ,其中 6人Rh表型为DCCee、3人为DCcee ;另 16名Del个体为RHD+ /RHD-杂合型 ,Rh表型均为DCcee。结论　D阳性个体RHD+ /RHD+ 纯合型多见 ,弱D型个体以RHD+ /RHD-杂合型为主 ,在Del表型个体存在高比率的RHD+ /RHD+ 纯合型 ;携带无效等位基因的个体多为RHD+ /RHD-杂合型     

1例部分D表型献血者RhD抗原表位及分子机制研究

目的 研究1例RhD抗原表型为部分D的献血者RhD抗原和分子机制.方法 利用血清学方法对1例初筛为RhD阴性样本进行RhD阴性确认并进行CE分型;采用D-screen检测该样本RhD抗原表位.利用Sanger测序法对RHD基因的全部外显子测序.建立数字PCR鉴定RHD基因型的方法并分析该例样本RHD基因合子型.采用Ro...     

Molecular characterization of RhD variant phenotypes among blood donors: A study from the coastal region of India

BackgroundRhD expression varies with population and ethnicity. Accurate typing of RhD antigen among blood donors is important to prevent development of anti-D among recipients of blood transfusion. We aimed to screen blood donors for variant D phenotypes and accurately characterize them by genotyping.Material and methodsWe have done prospective study on blood donors by performing RhD typing using three different commercial monoclonal anti-D reagents by both column agglutination and conventional tube techniques. Samples that showed ambiguous results were further screened with the Bio-Rad Partial RhD typing kit. Minor phenotyping for C, c, E, e antigens was performed. Multiplex PCR and Sequencing of all RHD exons with Sanger’s sequencing was performed for molecular characterization of variant D.ResultsA total of 16,974 blood donors were screened during the study period. Among them, 31 (0.18 %) donors were found to have a RhD variant phenotype. The male to female ratio was 10:1. The presence of ‘C’ antigen was noted among all RhD variant samples. Serological typing identified two samples as DV phenotype and the rest could not be characterized. Molecular genotyping characterized 90.3 % of the samples as Indian specific weak D type 150 variants. Three samples were subjected to Sangers sequencing and showed wild type pattern.ConclusionThe present study showed that the most common variant in this population was Weak D type 150. This study highlights that serological methods may serve as a screening tool, however, molecular techniques are essential for characterization of RhD variants.     

临沂地区RhD初筛阴性献血人群分子遗传背景研究

目的通过对临沂地区RhD初筛阴性献血者的研究,了解其分子遗传背景。方法 2011年1月～10月采集无重复54 458人份全血,其中盐水法初筛RhD阴性标本241份。对初筛RhD阴性标本运用本室建立的SSP-PCR作基因检测分型,并对未确定型别携带RHD的标本进行基因测序。结果临沂地区献血人群中RhD阴性比例为0.44%,241例初筛RhD阴性标本检测到1～10外显子全缺失169例,2～9外显子缺失22例,1227GA突变38例,845GA突变5例,3～6外显子缺失5例,RHD(711DelC)1例,DVa(Hus)1例。分子生物学试验确定的Del型共38例,占初筛RhD阴性的15.77%(38/241)。结论临沂地区RhD初筛阴性献血人群中RHD基因存在多态性,其中RHD1227A等位基因为临沂地区Del型的遗传标志性基因。     

Molecular background of D-negative phenotype in the Tunisian population

Background: Most studies of the molecular basis of Rhesus D‐negative phenotype have been conducted in Caucasian and African populations. A comprehensive survey of RHD alleles was lacking in people from North Africa (Tunisians, Moroccans and Algerians) which could be very efficient for managing donors and patients carrying an RHD molecular variant. We analyse the molecular background of D‐negative population in Tunisia in the present study. Materials and methods: Blood samples were collected from native Tunisians. A total of 448 D‐negative donors from different regions of Tunisia were analysed by RHD genotyping according to an adopted strategy using real‐time PCR, ASP‐PCR and sequencing. Results: Among the 448 D‐negative samples, 443 were phenotyped unequivocally as true D‐negative including three molecular backgrounds which were RHD gene deletion (n = 437), RHDψ pseudogene (n = 2) and RHD‐CE‐D hybrid gene (n = 4) with the respective frequencies of 0·9900, 0·0023 and 0·0046. The remaining five samples, in discordance with the serological results, were identified as two weak D type 11, one weak D type 29, one weak D type 4·0 and one DBT‐1 partial D. Conclusion: This study showed that the Tunisian population gets closer to Caucasians, given that the RHD gene deletion is the most prevalent cause of D‐negative phenotype, but it is slightly different by the presence of the RHDψ pseudogene which was found with a very low frequency compared with that described in the African population. Nevertheless, the relative occurrence of weak D variants among studied serologically D‐negative samples make necessary the adaptation of RHD genotyping strategy to the spectrum of prevalent alleles.