HbF变异体对三种不同HbAlc检测系统的干扰评价

目的评价血红蛋白变异体F(HbF)对三种糖化血红蛋白(HbA1c)检测系统测定结果的干扰。三种方法分别为离子交换高效液相色谱法(IE-HPLC)、硼酸盐亲和层析高效液相色谱法(AC-HPLC)和免疫抑制比浊法。方法分别用三种检测系统检测血红蛋白结构正常标本及含有胎儿血红蛋白(HbF)的标本,依据美国国家糖化血红蛋白标准化计划(NGSP)的判定标准,对检测结果进行比对分析和偏倚评估。结果以Primus Ultra2(AC-HPLC)为比较系统,IE-HPLC和免疫比浊法为检测系统,两种检测系统与比较系统检测正常样品HbA1c值的相关性良好。当HbF≤8.75%时,IE-HPLC测定结果与比较系统偏差6%;HbF为17.50%时,IE-HPLC测定结果与比较系统偏差6%;当HbF浓度达35.00%~70.00%时IE-HPLC无法检测出结果。免疫比浊法测定不同HbF浓度HbA1c值与比较系统偏差均6%,测试几乎不受HbF的干扰。结论 HbF对不同HbA1c检测系统的干扰程度不同,临床实验室在进行HbA1c检测时,应注意血红蛋白变异体的存在,必要时选用替代指标或者合适的方法进行HbA1c测定以防止干扰的发生。     

HbF变异体对三种不同HbA1c检测系统的干扰评价

目的 评价血红蛋白变异体F(HbF)对三种糖化血红蛋白(HbA1c)检测系统测定结果的干扰。三种方法分别为离子交换高效液相色谱法(IE-HPLC)、硼酸盐亲和层析高效液相色谱法(AC-HPLC)和免疫抑制比浊法。方法 分别用三种检测系统检测血红蛋白结构正常标本及含有胎儿血红蛋白(HbF)的标本,依据美国国家糖化血红蛋白标准化计划(NGSP)的判定标准,对检测结果进行比对分析和偏倚评估。结果 以Primus Ultra2(AC-HPLC)为比较系统,IE-HPLC和免疫比浊法为检测系统,两种检测系统与比较系统检测正常样品HbA1c值的相关性良好。当HbF≤8.75%时,IE-HPLC测定结果与比较系统偏差<6%; HbF为17.50%时,IE-HPLC测定结果与比较系统偏差>6%; 当HbF浓度达35.00%～70.00%时IE-HPLC无法检测出结果。免疫比浊法测定不同HbF浓度HbA1c值与比较系统偏差均<6%,测试几乎不受HbF的干扰。结论 HbF对不同HbA1c检测系统的干扰程度不同,临床实验室在进行HbA1c检测时,应注意血红蛋白变异体的存在,必要时选用替代指标或者合适的方法进行HbA1c测定以防止干扰的发生。     

变异血红蛋白对2种基于HPLC原理的糖化血红蛋白检测系统的影响

目的 比较亲和层析-高效液相色谱(AC-HPLC)系统和离子交换-高效液相色谱(IE-HPLC)系统测定血红蛋白(Hb)结构不同的血液样本中HbAlc的结果 ,评价变异Hb对2种检测系统的影响.方法 分别用2种HPLC系统同时检测HbA1c含量在5%～11%、Hb结构正常的血液样本和含有不同浓度胎儿Hb(HbF)的血液样本以及α-地中海贫血(α-MA)和β-地中海贫血(β-MA)患者血液样本中的HbA1c浓度.结果 对于Hb结构正常、HbA1c含量在5%～11%的血液样本,2种HPLC检测系统的测定结果 差异无统计学意义(P>0.05).AC-HPLC法不受血液样本中HbF的干扰;若血液样本中HbF的含量超过8.8%,IE-HPLC法测定结果 存在一定的偏倚,当HbF含量达70% 时甚至无法测出结果 .对于α-MA和轻型β-MA患者的血液样本,IE-HPLC法的结果 明显高于AC-HPLC法(P<0.01、P<0.05),而重型β-MA患者的样本用IE-HPLC法甚至无法测得结果 .结论 用IE-HPLC法检测含变异Hb、HbF的样本中的HbA1c,结果 可能受干扰;而用AC-HPLC法则不受干扰.     

四种方法检测糖化血红蛋白的干扰性能评价

摘要：目的：探讨亲和层析法、高效液相色谱法（HPLC）、免疫比浊法和酶法检测糖化血红蛋白A1c（HbA1c）对游离胆红素（F-Bil）、结合胆红素（C-Bil）、血红蛋白（Hb）和乳糜微粒（CM）的抗干扰能力。 方法：收集EDTA-K₂抗凝新鲜血,制备HbA1c 10.4%和6.2%的高、低浓度标本,对4种方法检测HbA1c进行干扰实验。 结果：4种干扰物对亲和层析法、HPLC法检测HbA1c无干扰。CM≥15 000 FTU时,免疫比浊法测定HbA1c有干扰。F-Bil、C-Bil≥121.3 μmol/L时,酶法检测高浓度HbA1c有干扰;F-Bil≥196.8 μmol/L、C-Bil≥121.3 μmol/L时,酶法检测低浓度HbA1c有干扰。 结论：酶法检测HbA1c抗胆红素干扰能力有待改进,应剔除黄疸标本。免疫比浊法则应避免高CM标本。     

胶乳凝集比浊法测定糖化血红蛋白的分析性能评价

目的 对胶乳凝集比浊法测定糖化血红蛋白(HbA1c)的分析性能进行评价.方法 用日立7600-120全自动生化分析仪对胶乳凝集比浊法测定HbA1c的分析性能进行检测.结果 试剂稳定性结果表明,新配制的试剂测定低值时到第3天才保持稳定,测定高值时试剂以每天平均0.1%的速度下降;胶乳凝集比浊法测定HbA1c批内CV为1.56%,批间CV为0.0%,天间CV为0.86%,总CV为1.59%;比对实验胶乳凝集比浊法和高效液相色谱(HPLC)法测定HbA1c呈良好的线性关系,Y=0.904X+0.239,r=0.9915;当胎儿血红蛋白F(HbF)≤16.2%时,对胶乳凝集比浊法和HPLC法测定的结果均无影响;线性范围为2%～15.8%;健康人HbA1c95%参考区间4.04%～5.69%,不同性别组比较差异无统计学意义(P＞0.05),年龄方面,以45岁为界,45岁以上中老年人的参考范围略高于45岁以下人群的参考范围(P＜0.05).结论 胶乳凝集比浊法测定HbA1c是一种适用于临床实验室开展的经济、有效、简单可行的方法.     

高血脂高胆红素对IE-HPLC法检测糖化血红蛋白干扰的评价

目的探讨高血脂和高胆红素对离子交换高效液相色谱法(IE-HPLC)检测糖化血红蛋白的干扰。方法将收集的新鲜EDTA-K2抗凝全血标本分成4组:对照组(HbA1c6.2%)、糖尿病组(HbA1c≥6.2%)、高血脂组(TG 3~20mmol/L)、高胆红素组(TBIL 21~549μmol/L)。分别用硼酸盐亲和层析高效液相色谱法(AC-HPLC)和IE-HPLC检测HbA1c。结果当HbA1c≤18.7%时IE-HPLC检测HbA1c相关系数r=0.993;95%置信区间(95%CI)为-0.71~0.89;偏差(%)为-5.8%~6.8%;差异无统计学意义(P=0.198)。当HbA1c16.3%时IE-HPLC检测HbA1c相关系数r=0.997;95%CI为-0.31~0.67;偏差(%)为-5.8%~4.3%;差异有统计学意义(P=0.000),结果无显著干扰;当%HbA1c为16.3~18.7时结果出现正偏倚。当TG≤20.78mmol/L时IE-HPLC检测HbA1c结果相关系数r=0.995;95%CI为-0.26~0.50;偏差(%)为-5.5%~5.8%;差异有统计学意义(P=0.000)结果无显著干扰;当TBIL≤549.3μmol/L对IE-HPLC检测HbA1c相关系数r=0.990;95%CI为-0.08~0.63;偏差(%)为-14%~4.1%;差异有统计学意义(P=0.000);当TBIL≤342.1μmol/L对IE-HPLC检测HbA1c相关系数r=0.994;95%CI为-0.09~0.50;偏差(%)为―5.5%~4.1%,结果无显著干扰;当TBIL为380.7~549.3μmol/L时结果出现负偏倚。结论实验数据表明IE-HPLC检测HbA1c有良好的抗脂血能力;当TBIL≤342.1μmol/L和HbA1c16.3%对检测HbA1c无明显干扰能满足一般临床检测需求;当HbA1c标本的TG≥20.78 mmol/L、TBIL≥342.1μmol/L、HbA1c≥16.3%临床工作者应结合患者本身情况选择更合适检测HbA1c的方法。     

TINIA检测HbA_(1c)的性能评价及和IE-HPLC在HbE患者中的应用比较

目的评价免疫抑制比浊法(TINIA)检测糖化血红蛋白(HbA1c)的分析性能,与离子交换高效液相色谱法(IE-HPLC)检测HbA1c进行比对和偏倚评估,并评估血红蛋白(Hb)变异体E(HbE)对TINIA和IE-HPLC的影响。方法参考美国临床与实验室标准化协会(CLSI)发布的方法学评价系列文件(EP文件)及有关文献,对TINIA检测HbA1c的精密度、正确度、分析测量范围、分析干扰以及生物参考区间进行评价和验证,对TINIA与IE-HPLC检测Hb结构正常的血液样本和含HbE的血液样本进行对比分析和偏倚评估,分析患者平均血糖的相关性。结果 TINIA检测HbA1c的批内精密度2.08%,总不精密度2.94%;在HbA1c浓度为4.4%~18.3%范围内,理论值与实测均值呈线性相关(r=0.999 4);Hb、游离胆红素(FBiL)、结合胆红素(CBiL)、乳糜(CH)浓度分别为9.9~49.6μmol/L、65.6~328.0μmol/L、65.6~328.0μmol/L和3 000~12 000 FTU时,差异百分比均5%;厂商提供的参考区间(4.8%~5.9%)适用于临床实验室。对于Hb结构正常的血液样本,TINIA和IEHPLC的HbA1c检测结果差异无统计学意义(P0.05),两种方法的相关性良好(r2=0.999 2)。对于HbE浓度在5.4%~54.7%范围内的血液样本,TINIA的平均偏倚为-4.1%~1.9%,均5%,几乎不受HbE的干扰,HbA1c检测结果与平均血糖的相关性较好(r=0.998 8);当HbE6.4%时IE-HPLC检测HbA1c的实测值与理论值平均偏倚为5.7%~278.7%,差异百分比均5%,测定结果与预期理论结果有明显差异;两种方法检测结果差异有统计学意义(P0.01)。结论 TINIA检测HbA1c的分析性能符合临床测定的性能要求,不受HbE的干扰。当HbE6.4%时,IE-HPLC法检测HbA1c会受干扰,出现假性增高的情况。     

变异血红蛋白对2种基于HPLC原理的糖化血红蛋白检测系统的影响

目的比较亲和层析-高效液相色谱(AC-HPLC)系统和离子交换-高效液相色谱(IE-HPLC)系统测定血红蛋白(Hb)结构不同的血液样本中HbAlc的结果,评价变异Hb对2种检测系统的影响。方法分别用2种HPLC系统同时检测HbA1c含量在5%~11%、Hb结构正常的血液样本和含有不同浓度胎儿Hb(HbF)的血液样本以及α-地中海贫血(α-MA)和β-地中海贫血(β-MA)患者血液样本中的HbA1c浓度。结果对于Hb结构正常、HbA1c含量在5%~11%的血液样本,2种HPLC检测系统的测定结果差异无统计学意义(P>0.05)。AC-HPLC法不受血液样本中HbF的干扰;若血液样本中HbF的含量超过8.8%,IE-HPLC法测定结果存在一定的偏倚,当HbF含量达70%时甚至无法测出结果。对于α-MA和轻型β-MA患者的血液样本,IE-HPLC法的结果明显高于AC-HPLC法(P<0.01、P<0.05),而重型β-MA患者的样本用IE-HPLC法甚至无法测得结果。结论用IE-HPLC法检测含变异Hb、HbF的样本中的HbA1c,结果可能受干扰;而用AC-HPLC法则不受干扰。     

两种方法检测糖化血红蛋白结果的对比分析

目的比较2种方法测定糖化血红蛋白(HbA1c)的结果之间的差异。方法采集糖尿病患者静脉全血标本,使用两种方法,即胶乳增强免疫比浊法在日立7170A全自动生化分析仪上,与高效液相色谱法(HPLC)在全自动糖化血红蛋白仪BIO-RADD10上,同时测定其HbA1c,对结果进行比较分析。结果 HPLC(X)与胶乳增强免疫比浊法(Y)测定结果呈明显正相关(r=0.9682);回归方程:y=1.127x-0.004。但胶乳增强免疫比浊法测定HbA1c的结果值较HPLC法明显偏高(P<0.01),以HPLC法为参照,胶乳增强免疫比浊法结果的相对偏倚为12.65%。结论实验室应尽量采用HPLC法为检测方法,或以其为参考方法对胶乳增强免疫比浊法进行校准或设置相应的参考区间,以提高结果的准确性与可靠性。     

糖化血红蛋白与空腹血糖联合检测的临床意义及其相关性探讨

目的探讨糖化血红蛋白(HbA1c)与空腹血糖(FGB)联合检测的临床意义及其相关性。方法选择2007年8月～2011年4月我院收治的120例糖尿病测定FBG和HbA1c水平,并分析HbA1c和FBG的关系。结果 120例糖尿病中HbA1c≤6.0%27例,FBG(5.97±1.46)mmol/L;HbA1c 6.1%～7.0%32例,FBG(7.89±2.62)mmol/L;HbA1c 7.1%～8.0%23例,FBG(9.25±2.98)mmol/L;HbA1c 8.1%～10.0%20例,FBG(10.83±3.46)mmol/L;HbA1c≥10.0%18例,FBG(12.25±4.05)mmol/L。其HbA1c和FBG相关性分析结果显示随着HbA1c水平增高,FBG也相应升高,二者呈正相关(r=0.8751,P<0.01)。结论 HbA1c联合FBG检测对糖尿病的诊断、治疗、转归和预后判定具有显著意义,应定期进行检测。     

Silent hemoglobin variants and determination of HbA(1c) with the high-resolution program of the HPLC HA-8160 hemoglobin analyzer

OBJECTIVES: Evaluation of HbA1c determination with an automated ion-exchange high-performance liquid chromatography (HPLC) method in patients with clinically silent hemoglobin (Hb) variants. DESIGN AND METHODS: HbA1c values were determined with the Arkray HA-8160 ion-exchange HPLC using the high-resolution, 4.2-min beta-thalassemia screening mode in patients with silent hemoglobin (Hb) variants, namely, Hb Graz, Hb Sherwood Forest, Hb O Padova, and HbD. RESULTS: All of these hemoglobin variants caused additional peaks in the chromatograms, without HbA1c results in patients with Hb Graz and Hb Sherwood Forest, and demonstrated extra peaks with HbA1c results that were clinically too low for patients with Hb O Padova and in the patient with HbD. CONCLUSIONS: The development of this automated HPLC method modification with high-resolution beta-thalassemia screening mode aids identification of interference due to some clinically silent Hb variants in HbA(1c) determination.     

Evaluation of HbA1c determination methods in patients with hemoglobinopathies

OBJECTIVE: To evaluate commercially available determination methods for HbA1c in patients with hemoglobin variants. RESEARCH DESIGN AND METHODS: HbA1c values were determined with various commercially available methods, including ion-exchange high-performance liquid chromatography (HPLC), boronate affinity assay, and immunoagglutination in patients with the hemoglobin mutations Hb Graz, Hb Sherwood Forest, Hb O Padova, Hb D, and Hb S. RESULTS: The effect of hemoglobinopathies on glycohemoglobin measurements was highly method dependent. The HPLC methods for HbA1c determination lacked the resolution necessary to differentiate hemoglobin variants. They demonstrated additional peaks in the chromatograms and HbA1c results either too low or too high compared with the nondiabetic reference range. With all immunoassays, Hb Graz demonstrated falsely low values. The other hemoglobinopathies in our study caused falsely low and/or high HbA1c results in immunoagglutination methods. The boronate affinity method showed values in an acceptable range for all hemoglobin variants. CONCLUSIONS: Because of the local occurrence of Hb variants and the ethnic origin of a given population, every individual laboratory must establish and validate its own assay method. In managing diabetic patients, knowledge of hemoglobinopathies influencing HbA1c determination methods is essential because hemoglobin variants could cause mismanagement of diabetes resulting from false HbA1c determinations.     

血红蛋白异常对高效液相色谱法检测糖化血红蛋白的影响

目的探讨血红蛋白（Hb）异常对高效液相色法谱检测糖化血红蛋白（HbA1c）的影响。方法使用国内常用的硼酸盐亲和层析高效液相色谱法（PDQ系统）及离子交换层析高效液相色谱法（D-10系统）进行含有不同（梯度）浓度HbF与正常人血液混合的标本的干扰实验及检测40例Hb异常的临床标本中的HbA1c。结果在干扰实验中,D-10系统在HbF在浓度超过40%时,可能无法获得检测结果,当HbF浓度在10%～40%时,结果高于实际值,HbF在浓度低于10%时,结果与实际值相似;PDQ系统在HbF浓度超过40%时,结果可能略低于实际值,HbF浓度低于40%时,可以获得好的结果。在40例Hb异常的临床标本中,D-10系统检测结果明显高于PDQ系统,差异具有统计学意义（P〈0.05）,其中有5例Hb异常总量超过35%的标本在D-10系统中无法获得检测结果,而PDQ检测获得可接受的结果;Hb异常总量低于10%时,两种方法检测结果有较好的一致性（P〉0.05）。结论硼酸盐亲和层析抗Hb异常的干扰能力优于离子交换层析。     

Evaluation of conditions associated with glycated hemoglobin values below the reference range

BACKGROUND: Evaluation of conditions associated with glycated hemoglobin (HbA1c) values below the reference range in HbA1c determinations. METHODS: Over a time period of 5 years, HbA1c results were determined with the ion-exchange high-performance liquid chromatography (HPLC) method HA-8140 Menarini. RESULTS: Of approximately 20 000 HbA1c results analyzed, 9 were below the reference range. The reason for HbA1c values below the reference range was found to be liver cirrhosis in 6 patients, anemia with hematological neoplasms in 2 patients, and elevated fetal hemoglobin > 1.5% in one patient. The silent hemoglobin (Hb) variant Hb Graz in 6 patients, Hb Sherwood Forest in 1 patient, homozygote HbS in one patient, and gross hypertriglyceridemia in one patient demonstrated no HbA1c result. CONCLUSIONS: In patients with liver cirrhosis, HbA1c measurements should be used with caution when evaluating long-term glucose control, and samples with suspected Hb variants should be analyzed by hemoglobin electrophoresis. Our study underscores the need for clinical laboratories and physicians to be aware of the limitations of their HbA1c assay methods as well as of the importance of visual inspection of ion-exchange chromatograms to detect HbA1c values below the reference range and abnormalities caused by the interference factors described here.     

Effect of HbE and HbH on HbA1C level by ionic exchange HPLC comparing to immunoturbidimetry.

BACKGROUND: The hemoglobin (Hb)A1C level is widely used to monitor diabetes mellitus patients. The N-terminal amino acid valine of its beta chain is glycated. The assay of HbA1C is based on differences in the charge, chemical and structural properties of the protein. METHODS: There are fully automated instruments available in clinical chemistry laboratory to assay HbA1C level. The effect of hemoglobinopathies was studied between an ionic exchange high-pressure liquid chromatography (HPLC) (Bio-Rad Laboratories, USA) and immunoturbidimetry (BM/Hitachi 912 with Roche HBA1CII, Germany-Japan) assay. The influence of high level HbF relative to the HbA1C level by ionic exchange HPLC is known. The effect of HbE and HbH to the HbA1C level by ionic exchange HPLC comparing to immunoturbimetry was examined. The evaluation was performed on 34 normal controls (A2A), 17 beta thalassemia traits (A2 upward arrow A), 36 HbE heterozygotes (EA), 37 HbE homozygotes (EE), 36 beta thalassemia/HbE (EF/EFA), 11 EABart's diseases (EABart's), 34 Hb H diseases (A2/CSAH) and 13 cord blood samples (FA). CONCLUSIONS: Hemoglobinopathies can impact on the assay of HbA1C level such as HbE and HbH to ionic exchange HPLC. Although not studied as yet, this effect may influence the other methods such as affinity chromatography.     

糖化血红蛋白不同组份的临床价值探讨

目的探讨糖化血红蛋白不同组份的临床价值。方法应用D-10糖化血红蛋白分析仪（高效液相色谱法）分析186例糖尿病（DM）患者全血中7种糖化血红蛋白组份[糖化血红蛋白A1亚组份（HbA1c、HbA1a、HbA1b）、抗碱血红蛋白（HbF）、不明蛋白组份（unknow）、蛋白组份3（P3）和不稳定糖化血红蛋白（LA1c_CHb-1）]水平。同时检测患者血糖（Glu）、C肽（C—P）、三酰甘油（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）、载脂蛋白A—I（apoA—I）、载脂蛋白B（apo B）和脂蛋白（a）[LP（a）]。结果Glu、HbA1a、HbA1b、HbF、P3和unknow与HbA1c呈正相关（P〈0．05），LA1c_CHb-1、C—P与HbA1c成负相关（P〈0．05）。186例DM患者HbA1c与血脂7项均无明显相关性，但其中101例男性患者HbA1c与总胆固醇（TC）呈正相关（r=0．333，P〈0．05）。结论除HbA1c外，HbA1a、HbA1b、HbF和P3在DM疗效评估中具有重要作用。DM患者，尤其是男性患者，在控制血糖的同时应辅以降脂治疗。     

Capillary electrophoresis of hemoglobin.

Capillary electrophoresis (CE) has been used in a variety of in-house capillary isoelectric focusing (CIEF) and capillary zone electrophoresis (CZE) assays for the detection of hemoglobin (Hb) variants and the quantitation of HbA2 and HbF. A commercial kit has also been produced for the analysis of hemoglobin variants and thalassemia screening. Though CE methods have been shown to be able to detect many variants, final identification of the variant needs specialized testing such as DNA technology. Over the past 2 years, many instruments that had been used for these hemoglobin variant screening and thalassemia assays have been withdrawn from sale. Although CE HbA1c analysis is available, it cannot compete in turnaround time or cost with automated HPLC commercial instruments that give accurate HbA1c results in 3 or 4 minutes. Hence we do not anticipate a bright future for the analysis of hemoglobin by CE.     

Influence of iron deficiency anemia on glycated hemoglobin levels in non-diabetic Saudi women

ObjectiveStudies of patients with iron deficiency anemia (IDA) have shown a relationship between high glycated hemoglobin (HbA1c) and low hemoglobin (Hb) concentration. The present study was conducted to determine the influence of IDA on HbA1c in non-diabetic women.MethodsFifty-nine Saudi women (20 to 50 years old) were enrolled and categorized into groups according to their circulating hemoglobin concentration: Non-IDA (Hb ≥7.45 mmol/L; n = 38) and IDA (Hb ≤7.44 mmol/L; n = 21). The IDA group was further subdivided according to the severity of the IDA, as follows: mild (Hb 6.83 to 7.44 mmol/L; n = 9) and moderate–severe (Hb <6.83 mol/L; n = 12). HbA1c, Hb, ferritin, fasting blood glucose, and red blood cell (RBC) count were measured in each participant.ResultsHbA1c did not significantly differ between the groups, but the absolute HbA1c level was significantly lower in the mild and moderate–severe anemia groups than the non-anemic group, and was positively associated with Hb, ferritin, and RBC count. In addition, the HbA1c level was inversely associated with the Hb concentration.ConclusionsHbA1c is significantly associated with parameters related to IDA in non-diabetic Saudi women. Therefore, assessment of IDA-related parameters is recommended prior to making a diagnosis of diabetes.