α1,3半乳糖基转移酶新等位基因278C＞T的鉴定

目的研究中国汉族人群ABO血型系统中Bw变异型的分子遗传背景，发现并鉴定一个新的ABO等位基因。方法血型血清学方法鉴定1例ABO血型疑难样本，应用聚合酶链反应、逆转录-聚合酶链反应和DNA序列分析等方法对先证者ABO基因转录调控序列和全编码序列进行突变筛选和检测。结果血清学和家系调查鉴定该样本为Bw表型，对gDNA和cDNA研究发现该样本存在第6外显子278C／T杂合，单倍体分析发现一种新的Bw等位基因，该等位基因与B101相比，差异仅在第6外显子的278C〉T错义突变，导致多肽链P93L替换，该突变点位于以前报道的Bw等位基因功能域之外。结论在中国人群中发现一种新的导致Bw变异型的ABO等位基因。     

α1，3半乳糖基转移酶基因721C〉T突变导致Bw亚型

目的研究红细胞ABO血型系统Bw亚型的分子基础. 方法通过标准血型血清学方法明确鉴定2个家庭3例Bw亚型,PCR扩增Bw亚型ABO糖基转移酶基因的增强子、启动子和第1～7外显子及侧翼内含子序列,PCR产物经割胶纯化后直接测序.同时将第6和7外显子克隆到pcDNA3.1(-)质粒,转化DH5α后进行序列分析.采用序列特异性引物-聚合酶链反应方法证实测序所发现的突变.结果直接测序发现3例Bw亚型的基因型为B/O杂合,其中糖基转移酶基因的第261位G杂合缺失,第721位C/T杂合.克隆证实一条染色体上为正常的O等位基因,另一条染色体上B等位基因(α1,3半乳糖基转移酶基因)存在第721位C>T突变,导致多肽链Arg241Trp替换.序列特异性引物-聚合酶链反应检测140份随机样本未发现此突变. 结论α1,3半乳糖基转移酶基因第7外显子721C>T突变可能是Bw亚型分子遗传基础之一.     

三例罕见ABO变异型Bw亚型的基因鉴定及序列分析

目的对3例ABO变异型Bw亚型进行基因鉴定及序列分析。方法采用常规血清学检测ABO血型, 并应用Sanger测序进行ABO基因鉴定。结果 3例ABO血型鉴定均符合Bw亚型, 基因分型分别为ABO*Bw.11/0.01.02、ABO*Bw.12/0.01.01、ABO*Bw.34/A1.02, 测序发现分别存在c.695T>C变异致多肽链Leu232Pro替换, c.278C>T变异致多肽链Pro93Leu替换, c.889G>A变异致多肽链Glu297Lys替换。结论本研究分别发现了Bw11、Bw12、Bw34亚型各一例。基因检测可作为血清学结果的补充确定ABO血型的亚型。     

α1,3半乳糖基转移酶基因721C>T突变导致B_w亚型

目的　研究红细胞ABO血型系统Bw亚型的分子基础。方法　通过标准血型血清学方法明确鉴定2个家庭3例Bw亚型,PCR扩增Bw 亚型ABO糖基转移酶基因的增强子、启动子和第1～7外显子及侧翼内含子序列,PCR产物经割胶纯化后直接测序。同时将第6和7外显子克隆到pc DNA3.1(- )质粒,转化DH5α后进行序列分析。采用序列特异性引物-聚合酶链反应方法证实测序所发现的突变。结果　直接测序发现3例Bw亚型的基因型为B/ O杂合,其中糖基转移酶基因的第2 6 1位G杂合缺失,第72 1位C/ T杂合。克隆证实一条染色体上为正常的O等位基因,另一条染色体上B等位基因(α1,3半乳糖基转移酶基因)存在第72 1位C>T突变,导致多肽链Arg2 4 1Trp替换。序列特异性引物-聚合酶链反应检测14 0份随机样本未发现此突变。结论　α1,3半乳糖基转移酶基因第7外显子72 1C>T突变可能是Bw亚型分子遗传基础之一。     

ABO亚型ABw07一家系的分子生物学研究

目的 研究一个ABO亚型ABw07家系的分子机制.方法 用单克隆抗体检测先证者红细胞ABO血型抗原.标准A、B、O红细胞检测先证者血清中的ABO抗体,采用聚合酶链反应技术扩增先证者ABO基因的第6和7外显子序列,PCR产物经酶切后直接测序分析.同时PCR产物经TOPO TA克隆到质粒载体中获得单链,对所得克隆进行ABO基因双向测序分析.家系调查采集先证者父母和姐姐的标本进行血型血清学实验和ABO基因第6和7外显子直接测序分析.结果 先证者红细胞有A、B抗原,同时血清中存在抗B抗体.直接测序分析发现第261位无缺失,第297位A/G、467C/T、526C/G、657C/T、703G/A、796C/A、803G/C、930G/A、1055C/A、1096A/G杂合,可推断为A102Bw07基因型.克隆测序得到两个等位基因A102和Bw07.与B101相比,Bw07第1055位G→A,导致1个氨基酸改变:第352位氨基酸精氨酸变成谷氨酰胺.家系调查显示先证者Bw07基因从母亲遗传所得,母亲血液标本ABO血型血清学特性和测序分析结果与先证者完全一致.结论 α-1,3-半乳糖基转移酶基因(B基因)第1055位G→A突变导致产生Bw07表型,其血清中可含有抗B抗体.     

ABO血型正反定型不一致样本的分子机制及其临床输血分析

目的 分析ABO血型正反定型不一致样本的分子机制,为临床ABO血型正反不符样本的鉴定和输血提供参考.方法 收集ABO血型正反不符的样本,选取其中血清学反应格局相似的样本6份,应用血清学方法、序列特异性引物-聚合酶链反应、ABO基因直接测序及TA克隆单体型分析等方法分析其分子机制,对其进行分类,并回顾性调查临床输血情况.结果 导致该6份样本ABO血型正反不符情况有3种:抗原减弱(2份)、抗体减弱(3份)、ABO亚型(1份).遵循同型或相容性输注原则进行输血均取得满意效果.结论 ABO血型存在异质性,血清学反应格局相同的样本可由多种机制引起.     

ABO基因278C>T突变导致Bw12一例

目的 对1例ABO疑难血型样本进行血清型和基因型鉴定.方法 用血清学方法对标本进行ABO血清型检测,根据血清型结果,选取ABO基因亚型检测试剂盒,用聚合酶链反应-序列特异性引物法进行基因亚型鉴定,用直接测序法对ABO基因的第6、7外显子进行序列测定.结果 血清学结果显示,该样本的红细胞上具有高凝集强度的A抗原和中等凝集强度的B抗原,样本血浆中含有弱凝集强度的抗-B抗体,初步判定为ABw型;B亚型基因分型试剂的结果显示,该样本ABO基因为ABw12型;直接测序结果显示,该标本ABO基因第6外显子序列为278CT、297GA,第7外显子序列为467CT、526CG、657CT、703GA、796CA、803GC、930GA杂合,即在基因型A102/B101的基础上,该序列nt278位发生了C＞T杂合突变,经与血型抗原基因变异资料库的数据比对,确定突变的等位基因为Bw12,基因型为A102/Bw12.结论 中国人群中发现A102/Bw12基因型.     

Bw11亚型家系表现ABO等位基因竞争现象的研究

目的分析携带ABO*BW.11等位基因的家系成员ABO血清学和分子生物学特征。方法应用血清学方法检测先证者及其家系成员共9人的ABO血型表型。采用PCR方法扩增ABO基因第6、7外显子并对扩增产物直接测序,同时克隆测序先证者及其父亲的标本。结果血清学检测初步判断先证者及其弟弟为AB亚型,先证者的父亲及其两女儿为B亚型。克隆测序发现先证者ABO等位基因第7外显子在B101的基础上第695位碱基发生T>C变异,表明为ABO*BW.11等位基因。先证者的父亲、弟弟及其两个女儿均携带该变异等位基因。A基因与BW.11以及BW.11与O基因同时遗传时竞争现象存在明显差异。结论ABO基因c.695T>C变异可能会导致Bw11亚型存在等位基因竞争现象。分子生物学方法结合血清学方法有助于精准鉴定ABO疑难血型。     

一个中国汉族ABO血型B亚型家系中发现新的B等位基因

目的　研究中国汉族人群ABO血型中B亚型的分子遗传背景,发现并鉴定ABO新等位基因。方法　随机选择10个正常的B型志愿捐血者样本作对照,对6例血型血清学鉴定为B亚型的样本,采用序列特异性引物-聚合酶链反应方法、ABO基因第6及第7外显子PCR产物直接测序,进行基因定型;并对B等位基因的第6外显子、第6内含子及第7外显子进行基因克隆和单倍体测序分析。结果　2例血清学为Bx、BW的B亚型样本中,发现一个新的B等位基因。该等位基因与B1 0 1 等位基因相比,差异仅在第7外显子nt6 95位T>C突变。进一步对其中一个Bx 血型的个体进行家系调查,结果该家系的11人中,7人带有该新B等位基因。而其余的4例B亚型样本及10例对照样本,ABO基因的第6、7外显子未发现新的点突变。结论　首次发现6 95 T>C变异的新B等位基因,该等位基因nt6 95位由T转变为C,2 32位氨基酸由亮氨酸转变为脯氨酸,可导致糖基转移酶活性的降低,表明ABO基因的第2 32位氨基酸对决定糖基转移酶活性至关重要。     

α-1,3- N-乙酰半乳糖胺基转移酶基因新变异的家系研究

目的通过对一例ABO亚型家系血清学和基因序列分析,研究该家系中α-1,3-N-乙酰半乳糖胺基转移酶基因新变异位点的特征。方法收集先证者及其3名家系成员血液标本,血清学方法进行ABO表型检测,荧光PCR进行ABO血型基因分型。通过对先证者ABO基因全编码区直接测序及第6、7外显子克隆测序方法进行基因序列及单体型分析。结果先证者血型为AxB亚型,ABO血型基因分型为A/B。克隆测序结果显示A新等位基因在ABO*A1.02序列基础上存在第7外显子c.797_798 insT变异。家系调查发现,先证者及其姐姐新变异均遗传自其父亲。c.797_798insT新变异序列已注册基因数据库(MK125137)。结论α-1,3-N-乙酰半乳糖胺基转移酶基因第7外显子c.797_798insT新变异为可遗传变异,可导致A抗原表达减弱。     

浅谈新生儿溶血病

由于与其他新生儿常见病相比致死率较高,且容易造成新生儿终身与疾病相伴,所以新生儿溶血病一直以来都受到妇儿医疗学界的重视,而近年来随着医学技术的发展,新生儿溶血病的检测和临床治疗手段有了很大的进步,其痊愈率也在不断上升,关于对新生儿溶血病的认识和分析,本文将从发病机理、临床表现及检查、治疗这三个方面为切入点进行.     

一例ABO血型重组等位基因的分子特性

目的分析一例ABO血型重组等位基因的分子特性。方法ABO表型鉴定采用试管法。ABO基因和FUT1基因编码区序列检测采用PCR测序法。利用等位基因特异性引物扩增测序技术鉴别先证者ABO等位基因。先证者及其母亲ABO基因全长序列测定采用二代测序方法。结果先证者红细胞与抗H不凝集,FUT1基因为c.551_552del AG纯合,判定先证者为类孟买型。先证者ABO基因双链测序结果为c.261G/del、467C>T、c.526C>G、c.657C>T、c.703G>A、c.796C>A、c.803G>C、c.930G>A杂合。单链测序结果显示先证者有一个ABO*A1.02等位基因,另一个为ABO*O.01.01和ABO*B.01重组形成的等位基因。二代测序数据显示可能重组的位置在核苷酸c.375-269到c.526之间,家系分析显示先证者重组等位基因遗传自母亲。结论ABO血型等位基因存在重组现象。发现了1例ABO*O.01.01和ABO*B.01重组形成的新等位基因。     

Lower incidence of de novo donor-specific antibodies against HLA-DR in ABO-incompatible renal transplantation

Recently, in vitro experiments have demonstrated that anti-blood group A/B antibody binding to endothelial cells induce a protective effect against antibody-mediated injury. This study aimed to clarify the potential clinical benefit of ABO incompatibility in donor-specific HLA antibody (DSA)-induced chronic antibody-mediated rejection (ABMR). We enrolled 215 ABO-incompatible renal transplant (ABO-I) and 467 ABO-identical/compatible renal transplant recipients (ABO-Id/C). The prevalence of de novo DSA production and incidence of biopsy-proven chronic ABMR were compared between the two groups. The incidence of DR-associated de novo DSA was significantly lower in ABO-I than in ABO-Id/C (P = 0.028). Diagnostic biopsy for ABMR was conducted in 54 patients (11 ABO-I and 43 ABO-Id/C). Biopsy-proven chronic ABMR was lower in ABO-I than in ABO-Id/C (27.3% [3/11] vs. 44.2% [19/43]) patients. Our findings suggest that ABO incompatibility may cause low production of DR-associated de novo DSA, possibly resulting in a reduced incidence of chronic ABMR.     

新生儿ABO溶血病的护理体会

目的：研究新生儿的早期静脉丙种球蛋白联合间歇光疗治疗新生儿ABO溶血病的护理。方法对49例新生儿ABO溶血病患者随机分为两组,A组观察组25例,早期静脉注射丙种球蛋白联合间歇光疗,B组24例,对照组只接受间歇光疗。治疗后血清胆红素的交界处的临界值在A组低于B组(P<0.05)72h,A组皮肤黄疸住院时间,明显低于B组(P<0.05)。结论丙种球蛋白联合新生儿ABO溶血疗法,可迅速降低血清胆红素水平,缩短病程。     

ABO incompatible renal transplants and decreased likelihood for developing immune responses to HLA and kidney self-antigens

Immune responses to HLA and tissue-restricted self-antigens (SAgs) have been proposed to play a role in the pathogenesis of renal allograft (KTx) rejection. However, ABO incompatible (ABOi) KTx recipients (KTxR) following depletion of antibodies (Abs) to blood group antigens had fewer rejections. To determine the mechanisms, pre- and post-transplant sera from ABOi (n = 18) and ABO-compatible (ABOc) (n = 45) KTxR were analyzed for Abs against HLA class I and II by LABScreen single antigen assay. The development of Abs to SAgs was measured by ELISA. Immunity to Collagen IV (Col-IV) and cytokines induced were measured by ELISPOT. While 8/45 (18%) ABOc KTxR developed new donor specific antibodies to HLA (DSA) following transplantation, 0/18 ABOi KTxR developed DSA. ABOi KTxR failed to develop Abs to kidney SAgs (Col-IV and fibronectin (FN)). In contrast, 7 ABOc KTxR developed Abs to both Col-IV and FN. Col-IV stimulation of lymphocytes from ABOc KTxR demonstrated increased IFNγ, IL-17 and decreased IL-10. In contrast ABOi recipients following stimulation with antigens resulted in more IL10 and reduced IFN-γ and IL17 production. At one year, the GFR in ABOi KTxR were significantly better (p < 0.04) than ABOc KTxR. De novo DSA and immune responses to SAgs are reduced or absent in ABOi KTxR which we propose leads to less acute rejection and better long term function following ABOi KTx.     

Molecular genetic analysis for a novel Ael allele of the ABO blood group system

The ABO blood group is the most important system in clinical transfusion medicine. Previous studies on the genetic base of the common ABO group and some rare ABO subgroups have suggested that the molecular genetic background of the ABO gene in the Chinese population has specific character. In this study, we carried out a molecular genetic analysis of a family with an individual diagnosed as Ael subgroup by serological tests. A novel allele was identified in our A subgroup cases.     

ABO血型在肾脏移植病例中的分布

目的了解肾脏移植病例中ABO血型的分布关系. 方法将143例肾脏移植患者与一般人群ABO血型进行对比分析. 结果肾脏移植病例与一般人群的ABO血型分布差异无显著性.肾脏移植患者中血型分布依次为O型占40.5%、A型占32.9%、B型占18.9%、AB型占7.7%.虽然患者中B型(18.9%)低于一般人群(22.9%),O型(40.5%)高于一般人群(34.8%),但差异无显著性.结论 ABO血型与肾脏移植病例无明显关联.     

A prospective observational study to compare transfusion outcomes in ABO identical versus ABO non-identical single donor platelet concentrates: An experience from a tertiary healthcare center in India

ABO incompatible single donor platelet concentrates (SDPC) have a concern about unsatisfactory increments as well as possibility of hemolytic transfusion reaction. But from Indian population no study has commented on the clinical and laboratory outcome of ABO mismatched platelet transfusion. The aim of study was to compare transfusion outcomes in ABO identical versus ABO non-identical single donor platelet concentrates. In this prospective observational study, 400 SDPC transfusions among different patients were included. In group A (n = 200), ABO identical SDPC transfusions and in group B (n = 200) ABO non-identical SDPC transfusions were added. Corrective count increment (CCI), absolute count increment (ACI), percent platelet recovery (PPR) were calculated and incidents of hemolytic transfusion reactions were noted. In group A mean ± SD of ACI, CCI and PPR were as 30.78 ± 12.51, 15.10 ± 6.677, 39,948.9 ± 20,099.392. In group B, mean ± SD of ACI, CCI and PPR were – 25.4 ± 15.65, 12.509 ± 5.906, 33,559.2 ± 22,150.304. And when CCI, ACI, PPR were compared with group A and group B, statistically significant differences were noted (P < 0.05). There was statistically significant difference in CCI, ACI and PPR in oncology patients and other prophylactic recipients except patients with dengue and other infectious disease. But there was no hemolytic transfusion reaction noted in any group. Our study clearly establish the potential benefits of ABO-identical PLT transfusion. It also points out that in emergency conditions or when there is a paucity in inventory, ABO non-identical SDPC transfusion may be lifesaving and clinically significant.