深圳地区9569例妊娠期妇女血红蛋白电泳结果分析

目的：评估全自动毛细管血红蛋白（Hb）电泳技术在深圳地区妊娠妇女产前珠蛋白生成障碍性贫血（地贫）筛查及异常 Hb 检查的临床应用价值。方法采用 Sebia capillary2flex 型全自动毛细管电泳仪对深圳市龙岗区妇幼保健院产前门诊孕妇进行 Hb 电泳检测,分析 HbA2水平及异常 Hb 电泳条带及水平。结果检出951例异常 Hb 电泳结果（异常率9．9％）包括 HbA2＞3．5％;HbA2＜2．5％;HbF ＞10％及其他异常 Hb（包括 HbCS 、HbE 、HbD 、Hb NewYork 和 Hb Q-Tailand 等）。结论全自动毛细管 Hb 电泳在地贫产前筛查及异常 Hb 检测中具有重要意义,可为进一步分子确诊及产前诊断提供指导。     

应用全自动蛋白电泳系统检查地中海贫血

目的　对全自动蛋白电泳系统筛查地中海贫血效果进行观察及评价。方法　用法国Sebia公司生产的HydrasysLC电泳仪进行血红蛋白电泳 ,用其配套的Hyrys扫描系统对HbA、HbA2、HbF、HbH、HbBart’s、HbCS区带进行扫描 ,得出各种血红蛋白含量 ,用pH8.6醋酸纤维薄膜电泳作HbH、HbBart’s、HbA2含量对比 ,用微量血红蛋白电泳作血红蛋白区带对比 ;用碱变试验作HbF对比。结果　测定标本 2 4 72份 ,检出β地中海贫血 173例 ;α地中海贫血 :用pH8.6醋酸纤维薄膜电泳法检出HbH 6 8例、HbBart’s2 5例 ,用全自动蛋白电泳系统电泳方法检出HbH 6 3例、HbBart’s 2 2例 ,HbCS两种方法均为 13例。结论　全自动蛋白电泳系统能很好地分辨出增高的HbA2和HbF ,对β地中海贫血检查有良好的效果 ;在α地中海贫血的检查中 ,对HbCS的检出效果较好 ,并且能作含量测定 ,但HbH区带多较淡染 ,有些标本出现HbBart’s区带浓于HbH区带的现象 ,有假阴性结果出现。     

全自动毛细管电泳检测11519例血红蛋白的结果分析

目的分析全自动毛细管电泳的血红蛋白电泳检测结果,并探讨血红蛋白电泳对血红蛋白病的筛查价值。方法收集2011年1月至2012年6月来自该院和外院1岁以上进行血红蛋白检测的患者资料,记录通过毛细管电泳检测的血红蛋白结果及基因检测结果,分析检出率和基因符合率。结果在此期间共检测11 519例血红蛋白电泳标本,筛查出1 193例疑似血红蛋白病患者,检出率10.36%,其中疑似α珠蛋白生成障碍性贫血(地贫)患者395例,占3.43%,疑似β地贫患者755例,占6.55%。HbE变异体检出18例,占0.16%,HbJ-K变异体检出20例,占0.17%,HbN带1例,HbD带4例。α地贫基因符合率75.07%,β地贫基因符合率达97.37%。HbE与基因符合率100%。结论毛细管电泳仪可有效地分离血红蛋白各组分,血红蛋白电泳可以较好的筛查血红蛋白病。     

2012至2016年广西血红蛋白组分室间质量评价结果的回顾性分析

摘要：目的：评价实验室对血红蛋白组分中血红蛋白A2（HbA2）和血红蛋白F（HbF）的检测能力。 方法：2012至2016年每年进行2次有关HbA2和HbF的室间质量评价,每次评价5个样本;按室间质量评价流程要求,各实验室在规定时间内检测样本并上传检测结果,依据回报结果统计各检测系统5年的使用分布情况、各实验室的合格率,计算并分析各检测系统、各检测方法的离散程度及不同浓度水平质控品检测结果变异情况。 结果：2016年,高效液相色谱法（HPLC）和毛细管电泳法（CE）使用率分别提高至46.1%（82/178）、18.0%（32/178）;2012至2016年,HbA2和HbF合格率由51.5%（34/66）、60.6%（40/66）分别提高至93.3%（166/178）、92.1%（164/178）;各检测系统的平均变异系数（CV）逐年缩小;Bio-Rad VariantⅡ和Sebia CAPILLARYS 2检测系统高、中、低值HbA2质控品检测结果平均CV较好,可控制在6.0%内;HPLC和CE可定量分析HbA2和HbF指标,总体检测能力优于琼脂糖凝胶电泳法。 结论：通过室间质量评价考核实验室检测HbA2和HbF能力,量化HbA2和HbF指标,为地中海贫血筛查和防治工作提供质量保证与数据支持     

血红蛋白S病17例临床与实验诊断分析

目的:总结17例血红蛋白S病(HbS)各种分型的实验数据,以提高临床实验室诊断各类型HbS的水平。方法:对所有病例行血常规检测并作Hb区带定量;采用红细胞镰变试验、红细胞高铁血红蛋白还原试验(MHb-RT)和葡萄糖-6-磷酸脱氢酶/6-磷酸葡萄糖酸脱氢酶(G6PD/6PGD)的直接比值法(紫外比值法)检测G6PD缺陷症。结果:所检测的17例HbS患者中新生儿4例,儿童和成人共13例。发现杂合子16例,纯合子1例,其中HbS合并α-Thal状态者4例。新生儿或成人患者与正常组比较,在血常规和Hb电泳中都有明显区别。17例HbS患者中有6例同时合并G6PD缺陷症。结论:HbS为正色素性贫血。Hb电泳区带定量是诊断HbS的重要方法,镰变试验是鉴别HbS与HbD的确诊试验。G6PD缺陷症在非洲裔HbS患者中有较高的发生率,若2种遗传病集于一身,将会加重贫血症状。     

BIO-RAD D-10血红蛋白检测仪在地中海贫血筛查中的应用

目的:评价BIO-RADD-10血红蛋白(Hb)检测仪在地中海贫血筛查中的应用。方法:对BIO-RADD-10血红蛋白检测仪进行性能评价,并测定我院门诊及住院病人HbA2及HbF含量,同时采用法国Sebia全自动电泳仪进行血红蛋白琼脂糖电泳,比较两种方法对HbA2及HbF含量的检测结果。结果:BIORADD-10血红蛋白检测仪检测HbA2及HbF批内精密度为4.8%、2.77%和1.42%、1.7%,批间精密度为4.91%、3.97%和2.87%、2.73%。共检测了1026例临床标本,并通过全自动琼脂糖凝胶电泳进行定量扫描,得出BIO-RADD-10测定灵敏度为HbF88.3%、HbA297.7%,特异性为HbF96.7%、HbA2为95.6%,阳性预测值为HbF97.4%、HbA296.9%,阴性预测值为HbF85.4%、HbA296.6%。但如果有其他异常血红蛋白条带或血红蛋白H、Bart’s,D-10血红蛋白检测仪不能识别,只能分辨出未知峰。结论:BIO-RADD-10血红蛋白检测仪能够分辨出HbA2及HbF异常增高者,为β地中海贫血的初筛提供快速的诊断依据。     

毛细管电泳在β地中海贫血中的应用研究

目的：研究全自动毛细管电泳仪检测血红蛋白A2(HbA2)在β地中海贫血筛查中的临床应用价值。方法986例需进行地贫筛查者,分别采用全自动毛细管电泳和琼脂糖凝胶电泳进行血红蛋白A2检测,并与基因分析结果进行对比,分析两种方法的灵敏度和特异度。结果以HbA2≥4.8％,毛细管电泳诊断β地贫灵敏度和特异度分别为82.3%和87.6%,琼脂糖凝胶电泳诊断β地贫灵敏度和特异度分别为77.6%和81.7%,毛细管电泳对β地中海贫血诊断的灵敏度和特异度较高。结论毛细管电泳比琼脂糖电泳能更准确区分HbA2、HbF等条带,对β地贫筛查效果更佳。     

全自动血红蛋白电泳在诊断地中海贫血中的应用

目的:探讨全自动血红蛋白电泳在诊断地中海贫血中的价值.方法:使用法国Hydrasyscs电泳系统进行血红蛋白电泳检测地中海贫血,同时采用HbF碱变性试验对同一标本进行HbF含量测定,比较两种方法的检测结果.结果:351例标本中,应用全自动血红蛋白电泳检出α地中海贫血表型阳性19例,阳性率为5.26%;β地中海贫血表型阳性65例,阳性率为18.52%.全自动血红蛋白电泳可以分辨出HbF区带,并且能进行定量扫描,与HbF碱变性试验测得HbF含量基本一致.结论:全自动血红蛋白电泳电泳区带清晰,扫描定量准确,可以为地中海贫血的诊断提供重要依据.     

80例贫血患者血红蛋白电泳分析

目的　通过对 80例非急性失血性贫血患者血红蛋白电泳分析 ,探讨血红蛋白电泳在不同贫血病例中的意义。方法　采用法国sebia公司HYDRASYS全自动电泳系统对 80例非急性失血性贫血患者 (Hb :37～ 93g/L ,平均 6 8g/L)进行血红蛋白电泳 ,测定HbA2 和HbF值 ,并用BECKMAN COULTER公司Gen’S血球计数仪测定各标本的血液学参数。结果　非缺铁性小细胞性贫血患者与缺铁性贫血组、正常组比较HbA2 明显增高 ,HbF正常或增高 ,MCV值与正常组比较明显减低 ,白血病患者与正常组比较HbF值可增高。结论　血红蛋白电泳在地中海贫血筛查中有极其重要的作用 ,且在其它血液疾病的鉴别诊断、疗效判断和预后分析等方面也具有一定的作用     

毛细管电泳在HbCS-H病中的诊断价值

目的探讨毛细管电泳分析仪在诊断血红蛋白（Hh）cs—H病中的应用价值。方法用SebiaCapillar—ys2型毛细管电泳仪分别对经地中海贫血基因分析确诊的34例HbCS—H病患者和70例非HbCS—H病患者（包括42例-SEA／-0-a、19例-SEA／-a4.2、7例HbWS—H、2例HhQs—H）进行Hb电泳。采用缺口聚合酶链反应（Gap—PCR）和反向斑点杂交的方法对2组患者进行地中海贫血基因分析。结果毛细管电泳分析HbCS—H组的HbCS带含量为（1．89±1．33）％,HbH带含量为（1．07±0．86）％,HbA,带含量为（1．17±0．68）％;非HbCS—H组均未能检测到HbCS带,有部分标本未检出HbH带,HbA：带含量为（1．81±1．21）％。2组HbA：带含量比较差异有统计学意义（P〈0．05）。毛细管电泳仪诊断HbCS—H病的敏感性和特异性分别为88．2％和100．0％,阳性预测值为100．0％,阴性预测值为94．6％,诊断效率为96．2％,与地中海贫血基因分析比较差异无统计学意义（P=0．134）。结论毛细管电泳可以用来快速诊断HbCS—H病,能够为患者减轻部分经济负担。     

Comparison of two methods for the quantification and identification of hemoglobin variants

BackgroundThe Sebia Capillarys 2, a capillary electrophoresis method, was compared to a high performance liquid chromatography (HPLC) and electrophoresis at alkaline and acid pH for the presumptive identification and quantitation of hemoglobin variants.MethodsThe concordance of hemoglobin variant identification on the Sebia Capillarys 2 with the combination of HPLC and electrophoresis at alkaline and acid pH was evaluated by analyzing samples on both systems. The quantification, expressed as % of total hemoglobin, of the common hemoglobin variants on the Capillarys 2 and the Bio Rad VARIANT II β thalassemia methods were compared.ResultsThe % hemoglobin variant results for the two methods were similar for HbS and slightly different for HbC and D Punjab and significantly different for HbE. The Sebia Capillarys 2 correctly identified a number of hemoglobin variants.ConclusionThe Sebia Capillarys 2 is suitable for the presumptive identification and quantification of hemoglobin variants producing results comparable with existing HPLC and electrophoresis methods.     

Evaluation of a new Sebia kit for analysis of hemoglobin fractions and variants on the Capillarys system.

We evaluated the analytical performances of the new Sebia kit for quantification of hemoglobin fractions (HbA, HbF and HbA2) and structural hemoglobin variants on the Capillarys system. This automated capillary zone electrophoresis method uses an alkaline buffer with silica capillaries and spectrophotometric detection. Specimen stability was evaluated during 1 month. The reproducibility of migration and the imprecision of quantification were also investigated. Comparison with the Beckman P/ACE system was performed on 202 samples. A total of 131 subjects without any hematological abnormality were analyzed to establish the HbA2 reference ranges based on our local population. Quantification of the Hb fractions and variants exhibited excellent stability for 4 weeks of storage at 4 degrees C, with CVs < 0.3%. The imprecision of the migration normalized to that of HbA2 for all hemoglobins tested (fractions and variants) was low, with a CV of < 2.5%. At physiological and pathological levels, total imprecision ranged from 1.9% to 4.6% for HbA2, from 0.6% to 9.7% for HbF, and from 0.6% to 1% for HbS. Statistical analysis revealed a small proportional negative bias for HbA2 (-8.6%). Small systematic bias (-0.2%) and proportional bias (-28%) were observed for HbF. No statistically significant difference was found for HbS. The reference range for HbA2 was 2.1-3.2%. The Capillarys system is a fully automated and accurate system that gives high-resolution performance and displays appropriate characteristics for use as a routine method for the diagnosis of thalassemias and hemoglobinopathies.     

Newborn screening for hemoglobinopathies using capillary electrophoresis technology: Testing the Capillarys® Neonat Fast Hb device

ObjectivesTo diagnose hemoglobinopathies in newborns by separating and measuring the Hb fractions on high throughput capillary electrophoresis. To test and validate the Capillarys Neonat Fast Hb device (Sebia) on fresh and dry blood samples.Design and methodsThe Hb fractions in 1.600 cord blood samples from the multi ethnic Dutch population were separated and measured. Further, the sensitivity, specificity and reproducibility of the device in detecting abnormalities and measuring the Hb fractions were estimated.ResultsThe apparatus separated all significant Hb fractions that should be detected during newborn screening (NBS) with 100% sensitivity. The reproducibility of the migrations guaranteed putative specificity for the few relevant frequent variants observed (HbS, C, and E). The estimation of the HbA and F fractions proved reliable using a well-designed integration mode.DiscussionDue to the limited number of samples no cases with sickle cell disease or β-thalassemia major were found in this cohort. However, the heterozygous state for the common variants associated with these diseases was clearly recognizable. The measurements were sufficiently precise to recognize sickle cell disease, β-thalassemia major and intermedia and to identify carriers including possible β-thalassemia. Therefore, Capillarys Neonat Fast Hb (Sebia) can be considered as a valid instrument for NBS of the Hemoglobinopathies on fresh and dry blood samples.     

全自动毛细管电泳仪在珠蛋白生成障碍性贫血筛查中的应用

目的评价法国Sebia公司Capillarys 2全自动毛细管电泳仪在珠蛋白生成障碍性贫血筛查中的应用。方法采集2011年8～12月经血分析(红细胞平均体积小于80fL,红细胞平均血红蛋白浓度小于26pg的8岁以上患者)筛选后的292份标本用法国Sebia公司Capillarys 2全自动毛细管电泳仪对入选标本进行血红蛋白(Hb)毛细管电泳、血清铁、铁蛋白检查,同时作α-及β-珠蛋白生成障碍性贫血基因检查。结果 (1)共检测了292例临床标本,符合α-珠蛋白生成障碍性贫血表型者36例,β-珠蛋白生成障碍性贫血表型者135例。同时出现α-及β-珠蛋白生成障碍性贫血表型者2例。(2)高压毛细管电泳法用于α-珠蛋白生成障碍性贫血诊断的灵敏度为86.11%,特异度为98.37%,准确度为95.60%,阳性预测值为93.94%,阴性预测值为96.03%。(3)高压毛细管电泳法用于β-珠蛋白生成障碍性贫血诊断的灵敏度为97.04%,特异度为98.35%,准确度为97.66%,阳性预测值为98.50%,阴性预测值为96.75%。HbA2值大于4.0%时,β-珠蛋白生成障碍性贫血基因法检测阳性率达100.00%。结论法国Sebia公司Capillarys 2全自动毛细血管电泳仪对Hb区带分辨及定量准确度高,可为Hb病(包括珠蛋白生成障碍性贫血及异常Hb病)的初筛提供快速的诊断依据。     

清远地区新生儿珠蛋白生成障碍性贫血筛查结果分析

目的分析清远地区新生儿珠蛋白生成障碍性贫血(简称地贫)筛查和基因诊断情况,为早期治疗和遗传咨询等干预措施及地贫防治政策的制定提供依据。方法应用法国Sebia Capillarys全自动毛细管电泳仪对2015年5月至2016年3月清远地区出生的新生儿进行滤纸干血斑血红蛋白成分分析,初筛阳性者召回抽静脉血应用裂口-聚合酶链式反应(Gap-PCR)和聚合酶链反应(PCR)结合反向点杂交技术进行地贫基因确诊。结果 1 056例初筛阳性者标本中共检出地贫908例,符合率为85.90%。α-地贫确诊符合率为95.05%(691/727),β地贫确诊符合率为65.00%(208/320),α-地贫筛查符合率显著高于β地贫,筛查为HbE的13例新生儿中共召回9例,全部确诊为HbE杂合子,筛查符合率100.00%。确诊为α-地贫650例,检出率为61.60%,其中静止型210例(19.90%)、标准型408例(38.60%)、血红蛋白H病32例(3.00%);检出β地贫204例,检出率为19.30%;α-地贫复合β地贫41例(3.90%),重型β地贫4例(0.35%),异常血红蛋白9例(0.85%)。结论通过新生儿滤纸干血斑进行血红蛋白成分分析可有效筛查出地贫患儿,值得推广应用,清远地区属于地贫高发区,在该地区开展新生儿地贫筛查很有必要,从而提高出生人口素质。     

False positive testing for sickle hemoglobin in a blood donor with mild erythrocytosis and hemoglobin Geldrop St. Anna

Solubility testing for sickle hemoglobin is commonly performed to identify blood suitable for patients with sickle cell disease. A 32-year-old Caucasian male blood donor’s unit screened positive for sickle hemoglobin via solublity testing (Streck). As the donor was considered low risk for being positive for hemoglobin S (HbS), he self-referred to hematology for further evaluation. Testing with hemoglobin electrophoresis revealed the patient to be negative for HbS; however, 42 % fetal hemoglobin (HbF) was noted. Since this was higher than typically seen in hereditary persistence of HbF, deoxyribonucleic acid (DNA) sequencing of hemoglobin (Hb) was ordered through a referral laboratory. Hb gene sequencing revealed the patient to be heterozygous for Hb Geldrop St. Anna, a rare Hb variant. This variant has previously been shown to migrate in the HbF region with alkaline electrophoresis. The workup demonstrated that the oxygen dissociation curve was left-shifted consistent with slightly increased oxygen affinity of this variant. The patient’s hematocrits (Hct) from his past donations were 53 % and 54 % about two years apart and his Hct at his hematology evaluation was 53 %. This report describes the first case of Hb Geldrop St. Anna in the United States and was associated with a false positive HbS screen. This Hb variant is considered to be benign and has an increased oxygen affinity that is associated with mild erythrocytosis. The donor was allowed to continue donating blood products.     

深圳地区血红蛋白G的分子特征与血液学表型

目的分析深圳地区异常血红蛋白G(HbG)病的分子特征和血液学表型,进一步认识异常血红蛋白病的类型,为地中海贫血筛查提供参考数据。方法收集我院进行毛细管电泳法检测的72397例样本,对检出异常血红蛋白者采用Sanger测序法进行DNA测序鉴定,并分析其红细胞参数,采用跨越断裂点聚合酶链反应(Gap-PCR)法和PCR结合反向点杂交(PCR-RDB)法检测并进行地贫基因分型。结果共检出6种HbG,均为杂合子,发生率为0.047%,位于α链异常的为Hb G-Honolulu和Hb G-Waimanalo;位于β链异常的为Hb G-Taipei、Hb G-Coushatta、Hb G-San José和Hb G-Siriraj。除1例Hb G-Honolulu合并β地贫(IVS-Ⅱ-654杂合突变)表现为轻型地贫特征,其余各类型HbG单纯杂合子的血液学表型均正常。新生儿Hb G-Taipei的毛细管电泳结果为HbA(8.6±2.4)%、HbF(89.1±3.1)%和Hb G-Taipei(2.3±0.6)%;Hb G-Coushatta结果为HbA 12.0%、HbF 83.7%和Hb G-Coushatta 4.1%。结论深圳地区HbG以杂合子多见,单纯HbG杂合子可表现为血常规正常,用毛细管电泳分析可发现携带者;合并地贫时可表现为地贫表型,临床应注意鉴别成人和新生儿HbG的毛细管电泳特征。     

Novel test method (sickle confirm) to differentiate sickle cell anemia from sickle cell trait for potential use in developing countries

The objective of this study was to develop a diagnostic testing method to detect HbS, distinguish sickle cell homozygotes from heterozygotes, and overcome testing barriers encountered in laboratories in underdeveloped countries. Blood samples positive and negative for sickle cell were subjected to the standard hemoglobin solubility test followed by a variety of centrifugation and filtration procedures. Each procedure was evaluated for the ability to remove insoluble HbS from the sample. The hemoglobin types that remain (HbA, HbA2 and HbF) were measured spectrophotometrically or estimated visually allowing samples to be categorized into three genotypes (AA, AS and SS) as confirmed by hemoglobin electrophoresis. De-identified EDTA blood samples were obtained from Saint Louis University and Cardinal Glennon Children's hospitals and tested in the Department of Clinical Laboratory Science at Saint Louis University. The main outcome measures were turbidity of the solubility solution; color of the supernatant and the material on the surface of the solution following centrifugation; precipitate trapped on the filter paper; absorbance of the filtrate; and hemoglobin electrophoresis patterns. Centrifugation and filtration successfully separated HbS from HbA/A2/F allowing for the differentiation of seven sickle cell homozygotes from sixteen heterozygotes with a sensitivity and specificity of 100%. This method has the potential to reliably distinguish homozygous from heterozygous sickle cell patients and it is fast, inexpensive, and simple. These characteristics make Sickle Confirm a desirable method in developing countries like Haiti and Africa where sickle cell anemia is prevalent and modern diagnostic methods like electrophoresis, HPLC and nucleic acid testing are impractical.     

Utilization of monoclonal-antibody-based assay (hemocardtm) in screening for and differentiating between genotypes of sickle cell disease and other hemoglobinopathies

Sickle cell disease covers a group of conditions in which pathology may be attributed to the presence of sickle hemoglobin (HbS). The identification of HbS and other variants including those in combination with HbS is commonly achieved by cellulose acetate electrophoresis at alkaline pH. Because many hemoglobin variants with similar charges have similar electrophoretic migration patterns, they are difficult to differentiate by electrophoresis. The HemoCard^TM assays address this concern through the use of monoclonal antibodies capable of specifically recognizing the unique amino acid sub-stitution in the variant hemoglobin. The panel of HemoCard monoclonal antibodies confirms the absence and presence of HbA, HbC, HbE, HbS, and other sickling hemoglobin variants. The combination of alkaline cellulose acetate electrophoresis and HemoCard assays allows the technologist to reach a final confirmation of both common and much less common sickle cell disease genotypes, combinations of HbS with other hemoglobins that ordinarily do not produce sickle cell disease, and other clinically important hemoglobinopathies including HbE/ß-thalassemia and hemoglobin C disease.