离子交换HPLC法和亲和层析HPLC法检测糖化血红蛋白结果的比较和分析

目的比较两种不同原理高效液相（HPLC）法测定HbA1c结果的一致性。方法采用Primus的PDQ亲和层析高效液相仪和Bio-Rad的VARIANT离子交换高效液相仪对87例2型糖尿病患者、10例透析的尿毒症患者和5例新生儿脐带血测定的糖化血红蛋白进行比较和分析。结果两种方法的线性试验中离子交换HPLC法的平均回复率为98．9％,亲和层析HPLC法平均回复率为100．9％;精密度试验离子交换HPLC法的批内CV〈1％,批间CV〈2％,亲和层析HPLC法批内和批间CV均〈2％;87例糖尿病患者检测结果比较没有显著性差异,r=0．901,P〈0．001;尿毒症患者HbA1c结果离子交换法均高于亲和层析法;亲和层析HPLC法能准确检测新生儿脐带血的HbA1c,而离子交换法不能测出。结论两种方法检测HbA1c准确度很高,都能真实的反映患者的糖化血红蛋白和血糖水平,但检测尿毒症患者和新生儿脐带血的糖化血红蛋白结果有很大差异。     

琼脂糖凝胶电泳法测定糖化血红蛋白

目的：用琼脂糖凝胶电泳法直接扫描测定糖化血红蛋白。方法：运用Sebia全自动琼脂糖凝胶电泳系统分离血红蛋白区带，直接扫描后测定糖化血红蛋白。结果：本法测定糖化血红蛋白批内CV为1．05％－3．69％，批间CV为2．26％－4．67％。与微柱法相比较两者相关良好（r=0.96)。结论：琼脂糖凝胶电泳法测定糖化血红蛋白快速、准确，适用于糖尿病控制的临床检查和研究。     

免疫凝集法检测糖化血红蛋白的应用评价

目的了解免疫凝集法检测糖化血红蛋白试剂盒测定结果的准确性、可靠性,并对其进行方法学评价,为临床在分析不同方法检测糖化血红蛋白结果时提供参考。方法参照美国临床实验室标准化委员会的方法学评价方案,与美国BIO-RADVARIANTⅡ糖化血红蛋白分析仪检测HbA1c(%)的结果进行对比,同时监测放置时间对其结果的影响。结果免疫凝集法线性良好,稀释变异可接受,线性范围为0～5.2g/L,最低可检出限为0.074g/L。日间CV=4.33%,批内CV=3.02%,批间CV=3.39%,总CV=4.51%。血浆中高浓度胆红素、三酰甘油和尿素干扰HbA1c的检测。全血标本4℃条件下放置2周后结果稳定;溶血标本4℃条件下放置6个月结果稳定。结论免疫凝集法检测HbA1c的线性范围、稀释变异均符合临床检测要求,最低可检出限度为0.074g/L,不精密度小于5%。与美国BIO-RAD VARIANTⅡ糖化血红蛋白分析仪检测结果比较,差异无统计学意义。可采用在4℃保存的溶血标本作为室内质控品。     

Capillarys 2 Flex Piercing 糖化血红蛋白检测系统性能验证

目的：评价 Capillarys 2 Flex Piercing 全自动毛细管电泳仪检测糖化血红蛋白(HbA1c)的检测性能。方法参照美国临床实验室标准化协会(CLSI)及厂家声明对 Capillarys 2 Flex Piercing 全自动毛细管电泳仪测定 HbA1c 的精密度、正确度、可报告范围、抗干扰能力、不同检测系统结果一致性进行验证。结果Capillarys 2 Flex Piercing 全自动毛细管电泳仪检测正常HbA1c 水平的批内、批间变异系数(CV)分别为0.80%、0.92%,病理 HbA1c 水平的批内、批间 CV 分别为1.10%、1.32%;正确度验证的偏差均小于厂家声明的5%;可报告范围验证的回归方程为 Y =1.002X +0.023(r 2=0.023,P =0.000),在4.4%~18.3%范围内的检测结果能达到线性要求;在 HbA1c 浓度分别在低、中、高水平时,干扰物总胆红素、脂血、血红蛋白对测定HbA1c 的偏差绝对值均未超过5%;与 Bio-Rad Variant Ⅱ糖化血红蛋白分析仪检测结果的一致性分析的回归方程为 Y =0.9877X -0.1344(r 2=0.9942,P =0.000),呈良好的线性关系;基于生物学变异的最佳允许总误差为2.31%。结论Capillar-ys 2 Flex Piercing 全自动毛细管电泳仪检测 HbA1c 的性能达到 CLSI 指南和厂家说明书的要求,可以满足临床检测需求。     

两种方法测定糖化血红蛋白结果的对比分析

目的比较免疫抑制比浊法和离子交换层析法测定糖化血红蛋白(HbA1c)的差异。方法使用两种方法同时检测51份常规样品,进行相关性的比较,结果采用相对偏差、t检验及回归方程进行统计分析。结果免疫比浊法和层析法测定结果相比,低值偏低0.5%、高值偏高3.1%;数据统计分析,t值分别为0.399、1.726、0.560,P0.05,差异无统计学意义;回归方程:Y=0.9437X+0.3774,r=0.9611;Y1=9.81,SE%=1.86%;Y2=15.48,SE%=3.27%。结论免疫抑制比浊法和离子交换层析法测定HbA1c结果相比差异无统计学意义,两种方法测定HbA1c结果的系统误差属临床可接受水平。     

胶乳增强免疫透射比浊法测定全血糖化血红蛋白A1c

目的建立胶乳增强免疫透射比浊法测定全血中的糖化血红蛋白A1c.方法用免疫透射比浊法测定糖化血红蛋白A1c,并对其重复性、准确性、线性范围、相关性实验和干扰实验加以评价.结果高、低浓度的糖化血红蛋白A1c批内CV为3.2%和4.1%;批间CV为4.6%和5.8%,总CV分别为6.7%和4.6%.线性范围为2%～15%;平均回收率分别为106.7%和102.4%.与微柱法测定结果比较具有良好的相关性(P>0.05).一定程度脂浊和黄疸对测定无明显干扰,抗坏血酸对测定无影响.结论胶乳增强免疫透射比浊法测定糖化血红蛋白A1c具有较好精密度和准确度,并且快速、灵敏和可靠,适合临床实验室常规开展.     

NycoCard ReaderⅡ检测仪测定糖化血红蛋白的临床评价

[目的]研究挪威产NycoCard ReaderⅡ多功能全定量特种蛋白金标检测仪测定糖化血红蛋白（HbAlc）的效果。[方法]采取58例糖尿病患者的抗凝全血，分别使用NycoCard ReaderⅡ分析仪和微柱离子交换法测定样本HbA1c含量，对两种测定方法之间的相关性和NycoCard ReaderⅡ分析仪测定糖化血红蛋白结果的精密度、批内变异、批间变异、干扰因素进行研究。[结果]两种测定方法无显著性差异；NycoCard ReaderⅡ分析仪检测糖化血红蛋白，批内CV值〈1．0％，批间CV值〈2．5％；高浓度的葡萄糖、乳糜、胆红素、对检测结果没有影响。[结论]NycoCard ReaderⅡ分析仪检测HbA1c准确性好、精密度高、干扰因素少、速度快，仪器及试剂成本低廉，适合临床开展。     

两种糖化血红蛋白测定方法的比较

目的 比较免疫抑制比浊法和离子交换层析法测定糖化血红蛋白(HbA1c)的差异.方法 使用免疫抑制比浊和离子交换层析两种不同的方法同时检测73份常规样品,进行相关性的比较,结果采用相对偏差、t检验及回归方程进行统计分析.结果 免疫比浊法和层析法测定结果相比,正常组偏低0.15%、异常组偏低0.08%、不分组偏低0.11%,数据统计分析,t值分别为1.29、0.16、0.36,P>0.05,差异无统计学意义;回归方程:Y=0.886 4X+0.782,r=0.804 9;Y=0.822 6X+1.553 8,r=0.951 9;Y=0.899 2X+0.811 5,r=0.970 2.结论 免疫抑制比浊法和离子交换层析测定HbA1c结果相比差异无统计学意义,两种方法测定HbA1c结果的系统误差属临床可接受水平.     

酶化学法测定糖化血红蛋白性能的验证

目的对酶化学法测定糖化血红蛋白进行方法学性能的验证。方法参考美国国家临床实验室标准化委员会系列文件和相关文献,结合工作实际对酶化学法测定糖化血红蛋白进行精密度、准确度的分析测量范围和生物参考区间等进行评价,并将实验结果与厂家提供的分析性能或公认的质量指标进行比较。结果批内变异系数(CV)0.87%~1.29%,批间CV1.74%~2.12%;在3.0%~16.0%范围内线性良好Y=1.010X-0.004,r=0.999 7,平均回收率101.37%;该方法与TOSOH G7离子交换高效液相层析法二组比较差异无统计学意义(Y=0.962 2X+0.045,r=0.994 0,P>0.05);分析测量范围(AMR)验证和生物参考区间验证结果均符合质量要求。结论酶化学法测定糖化血红蛋白主要分析性能符合质量目标要求,适合临床检验科应用。     

两种糖化血红蛋白分析方法的评价

目的对离子交换高压液相层析法与金标免疫渗滤法检测糖化血红蛋白进行方法学分析。方法用离子交换高压液相层析法(X)和金标免疫渗滤法(Y)测定糖化血红蛋白进行线性范围、回收率、精密度、干扰因素、相关性及参考范围的分析。结果离子交换高效液相色谱法(HPLC法)与金标免疫渗滤法(金标法)的线性范围分别为4.3%~17.0%和3.6%~12.5%,两种方法的回收率均为101.3%,批内、批间的变异系数(CV)均<5%;10.2%mmol/L三酰甘油、684μmol/L总胆红素和HbF对两种方法均无干扰。金标法测定糖化血红蛋白的结果较HPLC法明显偏低(P<0.01);回归方程Y=0.89X 0.24(r=0.94,P<0.01);HPLC法与金标法的参考范围分别为4.2%~6.7%和4.1%~6.4%。结论金标法与HPLC法相比,糖化血红蛋白检测结果明显偏低。     

Determination of glycosylated hemoglobin by affinity electrophoresis on agarose gel

We describe a method for electrophoretic determination of glycosylated hemoglobin (GHb) on agarose gel membrane. Clear separation of GHb from non-GHb is achieved after 30 min electrophoresis at 60 V using a sodium citrate buffer solution (34 mmol/l, pH 6.5) containing 0.6 g/l dextran sulfate. The results obtained by this method correlate well with those by agar gel electrophoresis (r = 0.98) and column chromatography (r = 0.96). However, unlike column chromatographic methods, GHb values obtained on agarose gel are not affected by fluctuations in ambient temperature (18-28 degrees C), changes in pH (6.2-6.6) and ionic strength of buffer solution (26-40 mmol/l), or variant hemoglobin S and C. Also, electrophoresis on agarose gel membrane eliminates the lengthy preparative steps required for cellulose acetate electrophoresis. We conclude that agarose gel electrophoresis is a simple method for quantitative determination of GHb.     

Interference of fetal hemoglobin and labile glycosylated hemoglobin with measurements of glycosylated hemoglobin

We examined the effect of fetal hemoglobin and labile glycosylated hemoglobin on a number of diverse methods used to measure glycosylated hemoglobin. Samples were supplemented with various amounts of cord blood to give proportions of fetal hemoglobin ranging from 1 to 20% of total hemoglobin concentration. Procedures in which the separation of hemoglobin A1 from the major hemoglobin A fraction is based on differences in ionic properties (cation-exchange chromatography and electrophoresis) are subject to interference by fetal hemoglobin, whereas procedures that base the quantitation on other properties (colorimetry and affinity column chromatography) are not. The same procedures that are affected by the presence of fetal hemoglobin are also subject to interference by labile glycosylated hemoglobin. We conclude that the affinity chromatographic and colorimetric methods may give a more nearly accurate determination of glycosylated hemoglobin.     

Use of fructosamine test in diabetic children

OBJECTIVE: The goal of this study was to assess the effect of glucose and the contribution of the aldimine component on the measurement of fructosamine, the relationship of serum fructosamine with glycosylated plasma proteins, as measured by a new high-performance liquid chromatography methodology (Glyc PP-HPLC) and by an affinity chromatography (Glyc PP), and the ability of serum fructosamine to assess acute, short-term (1-2 wk), and long-term (2-3 mo) glycemic control. RESEARCH DESIGN AND METHODS: The measurement of fructosamine was unaltered by the addition of up to 27.5 mM glucose or by the elimination of the aldimine component of serum specimens by dialysis. Fructosamine was generated in vitro by incubating serum aliquots. This generation was dependent on time, glucose concentration, and temperature. RESULTS: Fructosamine (n = 27) correlated well with Glyc PP (r = 0.76, P less than 0.01) and significantly less with Glyc PP-HPLC (r = 0.46, P less than 0.01). Although oral glucose ingestion increased serum glucose acutely by 200%, fructosamine was unchanged at each time interval. Improving glycemic control decreased the mean serum fructosamine concentration from 3.68 (baseline) to 3.28 mM (P less than 0.01) at 1 wk and to 3.13 mM (P less than 0.01) at 2 wk. HbA1c correlated with fructosamine (r = 0.59) and Glyc PP-HPLC (r = 0.47) but correlated best with Glyc PP (r = 0.83). CONCLUSIONS: These results indicate the fructosamine assay is unaltered by serum glucose, solely measures the ketoamine component, correlates well with glycosylated plasma proteins measured by aminophenylboronic acid column chromatography, is unaffected by acute changes of serum glucose, and may be used to monitor changes in glycemic control over a 1-wk interval.     

Accuracy of a filter paper method for measuring glycosylated hemoglobin.

OBJECTIVE--To compare glycosylated hemoglobin (GH) results obtained by filter paper fingerstick collection and mailed for assay by affinity chromatography with results from a venous sample assayed by ion-exchange chromatography (HbA1) in a local laboratory. RESEARCH DESIGN AND METHODS--Fifty-eight volunteer subjects with insulin-dependent diabetes mellitus (IDDM), aged 5-24 yr, included patients at a referral-based IDDM clinic and subjects in an ongoing research study. We obtained two blood samples from each subject. One was collected by fingerstick onto filter paper, the other by venipuncture into a vacutainer. We sent filter paper samples to the Diabetes Research Laboratory (Univ. of Missouri, Columbia, MO) for analysis. Vacutainer samples were sent to the Clinical Chemistry Department of the Clinical Laboratory, University of Colorado Health Sciences Center. RESULTS--Results were highly correlated (r = 0.89, P = 0.0001). Fifty-nine percent were classified identically when results were normalized to SD units and grouped to suggest levels of clinical concern. CONCLUSIONS--The filter paper method is a convenient, accurate measure of glycosylated hemoglobin in young people with IDDM. It should be considered a credible alternative research and clinical tool.     

免疫比浊法检测糖化血红蛋白A1c的性能评价

目的评价Roche公司第3代免疫比浊法检测糖化血红蛋白A1c(HbA1c)的性能。方法分析免疫比浊法检测HbA1c的精密度、线性,评价Hb、胎儿Hb(HbF)、三酰甘油(TG)、总胆红素(T-Bil)和不稳定HbA1c对HbA1c检测的影响、抗干扰能力及其与HPLC法测HbA1c结果的相关性;比较免疫比浊法检测HbA1c与高效液相色谱(HPLC)法的一致性。结果免疫比浊法检测HbA1c的变异系数(CV)均<2.5%。在4.7%～15.4%范围内,理论值与实测均值呈线性相关,r2值达0.999。Hb、HbF、T-Bil、TG水平分别在57～177 g/L、1.7%～11.7%、27～265μmol/L、3.2～9.85 mmol/L范围时,差异百分比均<5%;当HbF>13.5%时,差异百分比>5%。不稳定GHb(样本葡萄糖糖含量0～55.56 mmol/L)对HbA1c检测无影响。免疫比浊法(X)与高效液相色谱法(Y)相关性良好,Y=1.0188X-0.2271,r2=0.971 3。结论免疫比浊法检测HbA1c性能良好,适合临床应用。     

Evaluation of an affinity chromatographic procedure for the determination of glycosylated hemoglobin (HbA1)

An affinity chromatographic method for the determination of glycosylated hemoglobin (HbA1) was evaluated. The procedure was shown to be precise, the within- and between-assay coefficients of variation being less than 5%. It was also shown to correlate well with electrophoresis (r = 0.968) and ion-exchange chromatography (r = 0.916). An inverse relationship was shown to exist between increasing temperature and HbA1 levels measured by affinity chromatography. A statistically significant difference was found for samples run at 20 degrees C and 25 degrees C respectively, suggesting that the method should be run in a temperature-controlled environment. The affinity procedure was also shown not to be affected by the type of anticoagulant, the concentration of hemoglobin in the hemolysate, and acetylation.     

2008至2009年度上海市糖化血红蛋白项目室间质评结果分析

目的分析上海市2008~2009年度糖化血红蛋白（HbA1c）项目室间质评结果,为提高HbA1c检测质量提供依据。方法收集2008~2009年参加上海市临床检验中心HbA1c室间质评医院的4次调查结果,对结果进行统计分析。结果 2008~2009年上海市开展糖化血红蛋白室间质评的医院数从2008年122家增加至2009年178家,合格率从2008年96.7%增加到2009年98.9%。部分的调查结果离散度〉10%[2008年第1次的低压液相（LPLC）1号样本（10.8%）、免疫比浊法2份人新鲜全血样本（15.4%和15.9%）;2008年第2次的免疫比浊法2号样本（10.5%）;2009年第1次的微粒色谱法2号样本（10.3%）;2009年第2次的微粒色谱法2号样本（11.0%）和免疫比浊法1号样本（10.0%）]。经过质控管理,至2009年第2次调查,整体的平均变异系数（CV）有明显改进,人新鲜全血样本各检测系统的结果无明显差异,除微粒色谱法外,符合率达到100%。结论通过加强质控管理,开展HbA1c标准化工作,推动HbA1c检测结果的准确、可比,为临床提供可靠的诊疗依据。     

2型糖尿病患者动脉粥样硬化病变与血清CTRP3和CTRP9水平的相关性研究

目的探讨2型糖尿病(T2DM)患者血清 C1q/TNF 相关蛋白3(C1q tumor necrosis factor related protein 3,CTRP3),C1q/TNF相关蛋白9(C1q tumor necrosis factor related protein 9,CTRP9)与动脉粥样硬化病变(atherosclerosis,AS)的相关性。方法随机选取2014年1月~2015年1月陕西省人民医院内分泌科住院的成人新发T2DM患者196例,入选T2DM组,并根据颈动脉彩色多普勒超声结果将其分为糖尿病伴动脉粥样硬化组(AS组,71例)和单纯糖尿病组(非AS组,125例),另外随机选取同期健康体检者152例作为正常对照组,测定所有受试者的体重指数(BMI),腰围(WC),臀围(HIP),腰臀比(WHR),收缩压(SBP),舒张压(DBP),空腹血糖(FBG),餐后2h血糖(2hPBG),糖化血红蛋白(HBA1c),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),CTRP3及CTRP9。结果①T2DM组血清与正常对照组比较CTRP3(0.67±0.34 ng/ml vs 0.99±0.32 ng/ml),CTRP9(9.04±3.11 ng/ml vs 12.11±2.89 ng/ml),HDL-C(1.48±0.27 mmol/L vs 1.23±0.57 mmol/L),FBG(4.62±0.53 mmol/L vs 9.41±2.61 mmol/L),2hPBG(5.90±0.80 mmol/L vs 11.55±3.20 mmol/L),HBA1c(4.86±0.45 mmol/L vs 8.79±2.14 mmol/L),LDL-C(2.48±0.36 mmol/L vs 3.58±0.89 mmol/L),TC(3.64±1.10 mmol/L vs 5.77±0.97 mmol/L),TG(1.23±0.34 mmol/L vs 1.71±0.75 mmol/L)差异有统计学显著性意义(t=-13.069~5.88,P值均<0.01)。②AS组与非AS组比较CTRP3(0.67±0.34 ng/ml vs 0.99±0.32 ng/ml),CTRP9(9.04±3.11 ng/ml vs 12.11±2.89 ng/ml),年龄(55.82±8.80岁 vs 62.49±6.54岁)差异有统计学显著性意义(t/χ2=0.036~7.345,P值均<0.01),两组血清TC(5.61±0.90 mmol/L vs 6.05±1.02 mmol/L),LDL-C(3.44±0.80 mmol/L vs 3.83±1.01 mmol/L)差异亦有统计学显著性意义(t=-2.083~-2.197,P值均<0.05)。③相关性研究:CTRP3与WC(r=-0.932),DBP(r=-0.932),FBG(r=-0.856),TG(r=-0.728),TC(r=-0.920)呈负相关(P值均<0.01),CTRP9和年龄(r=-0.958)呈负相关(P值均<0.01),与HDL-C(r=0.860)呈正相关(P<0.01)。④多元逐步回归分析:WC,DBP,FBG是CTRP3的独立危险因子(P<0.01),年龄,HDL-C是CTRP9的独立危险因子(P<0.01)。结论CTRP3,CTRP9可能具有抗动脉粥样硬化作用,是糖尿病动脉粥样硬化的保护因子。     

糖化血红蛋白及糖化白蛋白在妊娠期糖尿病初筛中的价值

目的探讨糖化白蛋白(GA)和糖化血红蛋白(Hb A1c)检测在妊娠期糖尿病(GDM)筛查中的价值。方法选取妊娠22~28周的孕妇289例,其中血糖正常组202例、GDM组87例。分别采用己糖激酶法检测血糖、高压液相层析法检测Hb A1c,酶法检测GA。结果 GDM组Hb A1c为(5.20%±0.09%),明显高于血糖正常组(5.03%±0.02%,P0.01);GA为13.48%±0.28%,与血糖正常组(13.39%±0.09%)比较差异无统计学意义(P0.05)。Pearson相关分析显示血糖与Hb A1c呈正相关(r=0.203,P0.01),血糖与GA无相关性(r=0.114,P0.05),Hb A1c与GA无相关性(r=0.041,P0.05)。以血糖正常组Hb A1c第95%位值(5.50%)作为判断依据筛查GDM,特异性为94.55%,敏感性为86.21%。结论 Hb A1c相对于GA而言,是筛查GDM更好的指标。     

糖尿病肾病患者血清淀粉样蛋白A与血糖及血脂代谢的相关性

目的探讨血清淀粉样蛋白A(serum amyloid A,SAA)与糖尿病肾病患者血糖及血脂代谢的关系。方法 115例2型糖尿病患者依据24h尿微量白蛋白排泄率分为单纯糖尿病组37例,微量白蛋白尿组40例,临床白蛋白尿组38例,同期35名体检健康者为对照组,测定4组空腹血糖、总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、血肌酐、血尿素氮、糖化血红蛋白、血清SAA、24h尿微量白蛋白排泄率,并进行比较。结果单纯糖尿病组SAA、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平高于对照组(P<0.01);微量白蛋白尿组SAA、空腹血糖、糖化血红蛋白、总胆固醇水平高于单纯糖尿病组(P<0.05);临床白蛋白尿组糖化血红蛋白、总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、SAA、血肌酐、24h尿微量白蛋白排泄率、三酰甘油水平高于微量白蛋白尿组(P<0.05);SAA与24h尿微量白蛋白排泄率、空腹血糖、糖化血红蛋白、总胆固醇呈正相关(r=0.463,P<0.01;r=0.278,P<0.05;r=0.402,P<0.01;r=0.216,P<0.05),与高密度脂蛋白胆固醇呈负相关(r=-0.265,P<0.05)。结论 SAA与空腹血糖、糖化血红蛋白、高脂血症及尿微量白蛋白关系密切,SAA水平增高是糖尿病肾病进展的重要危险因素。