高密度脂蛋白胆固醇直接测定法与硫酸葡聚糖50—镁法比较

目的 评价高密度脂蛋白胆固醇（ＨＤＬ－Ｃ）直接测定法在临床应用的可行性，方法 将直接测定法与硫酸葡聚糖（ＤＳ５０）－氯化镁沉淀法进行比较，并分析其线性范围，准确度和干扰因素。结果 直接法与ＤＳ５０沉淀法相关良好，Ｙ＝０．９２７Ｘ＋０．２１４，ｒ＝０．９６４，ＨＤｌ－Ｃ浓度为２．４５ｍｍｏｌ／Ｌ范围内线性良好，ｒ＝０．９９９，ＨＤＬ－Ｃ浓度两组低，中，高值血清标本（０．８４，１．３１，２．３１；０．     

对国产高密度脂蛋白胆固醇直接法测定试剂的评价

评价国产慈城生化试剂厂生产的ＨＤＬ－Ｃ直接测定法试剂在临床应用的可行性。将慈城直接测定法试剂与葡聚糖（ＤＳ５０）－氯化镁沉淀法和磷钨酸（ＰＴＡ）－氯化镁沉淀法进行比较，并分析其特异性、线性范围、准确度和干扰因素。慈城试剂与ＤＳ５０沉淀法相关良好，ｒ＝０．９７１７、Ｙ＝１．０５９Ｘ－０．０５７，ｇｎ ＰＴＡｉｆ ｘｘｌｕ，ｒ＝０．９８５１、－１．０８８Ｘ－０．０９。在ＨＤＬ－Ｃ浓度为３．３４ｍｍｏｌ     

高密度脂蛋白胆固醇PEG—修饰酶直接测定法与DS—镁法和PTA—Mg^＋＋的实 …

评价高密度脂蛋白胆固醇（ＨＤＬ－Ｃ）ＰＥＧ－修饰酶直接测定法在临床应用的可行性。方法：将ＰＥＧ－修饰酶直接测定法与硫酸葡萄聚经镁沉淀法进行比较，分析线线性范围，准确度和干扰因素。结果：ＰＥＧ－修饰酶法与硫酸葡聚糖－氯化镁法相关性良好。Ｙ＝１．１２ｘ－０．０８，ｒ＝０．９７４，ＨＤＬ－Ｃ浓度为２．５５ｍｍｏｌ／Ｌ范围内线性良好，ｒ＝０．９９８；ＨＤＬ－Ｃ浓度在两组低，中、高血清标本０．８５、１．４５     

血清高密度脂蛋白胆固醇均相法测定试剂的研制

目的研制高密度脂蛋白胆固醇均相法测定的试剂。方法利用多聚体和聚阴离子在表面活性剂的作用下对脂蛋白进行选择性遮蔽,仅使高密度脂蛋白中的胆固醇参与胆固醇反应。结果研制的试剂（Ｙ）与推荐方法ＤＳ５０Ｍｇ（Ｘ１）和ＰＴＡＭｇ沉淀法（Ｘ２）相关良好,分别为Ｙ＝１．０４０Ｘ１－０．０６３,ｒ＝０．９８８３;Ｙ＝０．９４４Ｘ２＋０．０７３,ｒ＝０．９６５７。在ＨＤＬＣ浓度为２．８４ｍｍｏｌ／Ｌ以下线性良好,ｒ＝０．９９８２,ＨＤＬＣ低、中、高值血清标本的批内和批间（ＣＶ）值分别为２．０７％、１．８１％、０．８０％和２．７７％、１．９７％、１．７３％,特异性好,平均回收率为９８％,胆红素、血红蛋白等干扰不明显。结论ＨＤＬＣ均相法测定结果的准确性与精密度符合临床要求。     

聚乙二醇修饰酶法直接测定高密度脂蛋白胆固醇的评价

本文用德国Ｂｏｅｈｒｉｎｇｅｒ Ｍａｎｎｈｅｉｍ公司生产的高密度脂蛋白胆固醇试剂盒在Ｄｌｙｍｐｕｓ ＡＵ－５６０型生化仪上建立了直接法测定血清ＨＬＤ－Ｃ的程序，并对此方法作了初步评价。ＨＤＬ－Ｃ浓度在２．６ｍｍｏｌ／Ｌ范围内线性良好，ｒ＝０．９９９；两组低，中，高浓度的标本批内ＣＶ２．１％－３．３％，批间ＣＶ２．２％－３．６％，平均回收率为９７．５％，浓度为６．２ｍｍｏｌ／Ｌ甘油三酯，１０５．６μ     

高密度脂蛋白胆固醇的直接测定法

基于选择性抑制法的原理，通过对聚阴离子和表面活性剂的筛选实验，以及试剂配方和实验条件优化实验，建立了直接测定高密度脂蛋白胆固醇（ＨＤＬ－Ｃ）的方法。本法的线性范围达３．９ｍｍｏｌ／Ｌ，批内ＣＶ为１．４５％～２．０２％；总ＣＶＯ １．７１％～３．１４％，和ＴＰＡ法比较：Ｙ＝０．９９２３Ｘ＋０．０５２，ｒ＝０．０９８３４，ｎ＝６６。加入ＨＤＬ组份的回收率为１００．１％，本法的特异性好，加入ＬＤＬ－Ｃ达     

用8—羟基喹啉作掩蔽剂2—（8‘—羟基喹啉—5’—磺酸—7‘—偶氮）—变色 …

目的：建立２－（８‘－羟基喹啉－５’－磺酸－７’－偶氮）－变色酸（８Ｑ５ＳＡＣ）血清钙测定法。方法 以８Ｑ５ＳＡＣ作显色剂，８－羟基喹啉掩蔽镁，在三乙酸胺缓冲介质中以分光光度法测定血清钙。结果：方法学线性范围０－４．５ｍｍｏｌ／Ｌ，平均回收率１００．８５％，批内ＣＶ０．８４％￣１．２４％，批间ＣＶ１．５２％￣１．５６％，与ＯＣＰＣ法对照，ｒ＝０．９９５，Ｐ〉０．５，与ＭＴＢ法对照，ｒ＝０．９８９，     

甲基百里香酚兰法直接测定红细胞内镁的实验研究

本文介绍用甲基百里香酚兰法直接测定红细胞内镁的实验方法，并对有关实验条件进行了探讨。该法显示镁浓度在２～１６ｍｍｏｌ／Ｌ内线性良好，ｒ＝０．９９９６。平均回收率为１００．７％；批内ＣＶ＝１．０４％，批间ＣＶ＝５．０３％。作者测定了２０例健康成人红细胞内鲜深度度为１．２９０～２．０５０ｆｍｏｌ／ｃｅｌｌ（＾－Ｘ±２ＳＤ）。     

偶氮氯磷Ⅲ比色法测定血清钙的实验研究

本文对偶氮氯磷Ⅲ（ＣｈｌｏｒｏｐｈｏｓｐｈｏｎａｚｏⅢ）比色法测定血清钙的实验方法作了系统性研究。结果表明：显色剂中偶氮氯磷Ⅲ浓度以１２０μｍｏｌ／Ｌ为宜，反应最适ｐＨ为２．８～３．２，最佳检测波长为６２０ｎｍ，灵敏度为１ｍｍｏｌ／Ｌ＝０．１６４Ａ，线性为４．５ｍｍｏｌ／Ｌ，批内（ｎ＝２０）和批间（ｎ＝４０）ＣＶ分别为２．０８％和２．２６％，平均回收率为１００．２％。本法（Ｙ）与ＡＡＳ法（Ｘ１）及ＡｒｓｅｎａｚｏⅢ法（Ｘ２）结果比较：Ｙ＝０．９８５６Ｘ１＋０．０３７０、ｒ＝０．９９１２，Ｙ＝０．９５４０Ｘ２＋０．１３６５、ｒ＝０．９８８７．溶血、脂血、黄疸不干扰测定，当标本中Ｍｇ２＋、Ｃｕ２＋、Ｆｅ２＋、Ｚｎ２＋分别达５ｍｍｏｌ／Ｌ、２ｍｍｏｌ／Ｌ、３００μｍｏｌ／Ｌ、３００μｍｏｌ／Ｌ时也不影响测定结果。     

全血2,3—二磷酸甘油酸比色测定法

本文介绍了用磷酸甘油酸变位酶测定２，３－二磷酸甘油酸的比色法。探讨了该法的最适反应条件，最适ｐＨ７．５％￣７．６。线性范围可达６．０ｍｍｏｌ／Ｌ。三份不同浓度标本批内ＣＶ１．１％￣２．１％，批间ＣＶ２．５％￣３．９％。６０例南京地区健康成人２．３－二磷酸甘油酸含量为１．６￣２．７ｍｍｏｌ／Ｌ，平均回收率９４％，与Ｓｉｇｍａ试剂盒比较测定结果无显著性差异（Ｐ〉０．０５），相关性良好，ｒ＝０．９８２。     

Comparison of a new method for the direct and simultaneous assessment of LDL- and HDL-cholesterol with ultracentrifugation and established methods

BACKGROUND: Automated electrophoresis combined with enzymatic cholesterol staining might improve routine assessment of LDL- and HDL-cholesterol (LDLC and HDLC), as an alternative to the Friedewald equation and precipitation. A new method (Hydrasys; SEBIA) that adapts the cholesterol esterase/cholesterol oxidase reaction within urea-free gels was evaluated. METHODS: Fresh sera from 725 subjects (512 dyslipidemics) were analyzed by electrophoresis, in parallel with sequential ultracentrifugation, beta-quantification, calculation, and precipitation. RESULTS: Electrophoresis was linear up to 4 g/L cholesterol, with a detection limit of 0.042 g/L cholesterol/band. Within-run, between-run, between-batch, and between-operator imprecision (CVs) were 1.6%, 2.0%, 1.5%, and 2.7% for LDLC, and 3.9%, 4.3%, 5.5%, and 4.9% for HDLC, and remained unchanged up to 6.3 g/L plasma triglycerides (TGs). Precision decreased with very low HDLC (<0.25 g/L). Serum storage for 3-7 days at +4 or -80 degrees C did not interfere significantly with the assay. Agreement with beta-quantification was stable for LDLC up to 5.07 g/L (r = 0.94), even at TG concentrations >4 g/L (r = 0.91). Bias (2.88% +/- 12%) and total error (7.84%) were unchanged at TG concentrations up to 18.5 g/L. Electrophoresis predicted National Cholesterol Education Program cut-points with <0.04 g/L error, exactly and appropriately classified 79% and 96% of the subjects, and divided by 2.4 (all subjects) and 5.8 (TGs >1.5 g/L) the percentage of subjects underestimated by calculation. One-half of the patients with TGs >4 g/L had LDLC >1.30 g/L. For HDLC, correlation was better with precipitation (r = 0.87) than ultracentrifugation (r = 0.76). Error (-0.10% +/- 26%) increased when HDLC decreased (<0.35 g/L). Direct assessment of the LDLC/HDLC ratio detected 45% more high-risk subjects than the calculation/precipitation combination. CONCLUSIONS: Electrophoresis provides reliable quantification of LDLC, improving precision, accuracy, and concordance over calculation, particularly with increasing plasma TGs. Implementation of methods to detect low cholesterol concentrations could extend the applications for HDLC assessment.     

低密度脂蛋白胆固醇保护性试剂匀相测定法的临床评价

目的 对低密度脂蛋白胆固醇(LDT-C)保护性试剂匀相测定法进行临床评价。 方法 分析了保护性试剂匀相测定法的精密度、准确性、特异性和干扰因素.并随机选取了219份病人血清标本,比较分析用保护性试剂匀相测定法直接测定与Friedewald公式和Planella公式计算的LDL—C结果。 结果 保护性试剂匀相测定法具有较好的精密度(批内、批间CV和总CV均小于3％)。线性范围至10.4mmol/L,最低检测浓度为0.08mmol/L,平均同收率为101.2％:基本不受极低密度脂蛋白(VLDL)、高密度脂蛋白(HDL)和血红蛋白的影响。在TG<4.52mmol/L时,用匀相测定法与Friedewald公式和Planella公式的计算法结果之间相关性良好,两种公式计算法结果之间的也有较好相关性;而在TG>4.52mmoL/L时,匀相测定法与两种计算法之间的相关性差。结论 保护性试剂匀相测定法简便、快速、结果准确,易于自动分析,适合在临床实验室常规检测应用。     

A new liquid homogeneous assay for the determination of HDL-cholesterol. A comparison to precipitation with phosphotungstic acid/MgCl2 and a lyophilized homogeneous assay.

We evaluated a new ready to use liquid assay for the homogeneous determination of HDL-cholesterol (HDL-C; Merck, Darmstadt, Germany) in comparison to phosphotungstic acid precipitation and a homogeneous assay, based on sulfated alpha-cyclodextrin and polyethylene glycol-modified enzymes (Roche Diagnostics/Boehringer Mannheim, Germany). The new liquid homogeneous HDL-C assay had inter-assay coefficients' of variation of less than 2.1%. The method is linear up to at least 3.11 mmol/I HDL-C, but even at 4.40 mmol/I the deviation from the expected value is less than 5%. Spinking experiments with low density lipoproteins and very low density lipoproteins proved that the new assay was specific for high density lipoproteins up to cholesterol associated with low density lipoproteins (LDL-C) and very low density lipoproteins (VLDL)-triglyceride concentrations of 18.13 and 22.60 mmol/l, respectively. Free fatty acids above 2mmol/l did not interfere. Icteric samples with bilirubin concentrations between 170 and 400 micromol/l did not show any systematic deviation compared to the precipitation procedure. In addition, serum hemoglobin concentrations up to 7.0 mmol/l and ascorbic acid up to 3000 micromol/l did not interfere with the HDL-C assay. An intermethod comparison including 120 samples revealed good agreement of the liquid HDL-C assay and the precipitation procedure (y = 0.943x + 0.074 mmol/l; r = 0.992). The new homogeneous HDL-C assay is thus precise, comparable and robust. Due to its ease of handling this assay will significantly facilitate attempts to include the differentiation between HDL-C and LDL-C in the routine screening for cardiovascular risk factors and in the monitoring of lipid lowering therapy.     

Comparison of a homogeneous assay with a precipitation method for the measurement of HDL cholesterol in diabetic patients

OBJECTIVE: To compare direct-measured HDL cholesterol with HDL cholesterol measured by a precipitation method. RESEARCH DESIGN AND METHODS: We compared a homogeneous assay for direct HDL cholesterol analysis with the phosphotungstic acid magnesium chloride precipitation method in 55 type 1 diabetic patients, 70 type 2 diabetic patients, and 82 nondiabetic normal control subjects with plasma triglyceride levels <4.6 mmol/l. The cholesterol content of HDL determined by the direct assay was overall 0.1 mmol/l higher in all three groups than HDL cholesterol measured after precipitation, but the two methods were closely correlated (r(2) = 0.98, P < 0.001). RESULTS: HbA(1c), blood glucose, serum albumin, serum bilirubin, or triglyceride did not influence the differences of the two HDL cholesterol measurements. Because we have previously shown HDL cholesterol isolated by phosphotungstic acid precipitation to be lower than that by ultracentrifugation, the positive bias found in this study was expected. It seems that the direct HDL cholesterol assay reacts with apolipoprotein (apo) B-containing lipoproteins in the fraction with a density of >1.063; these apo B-containing lipoproteins are suggested to be coprecipitated with the phosphotungstic acid method. We also measured LDL cholesterol directly by a LDL cholesterol plus method and found no significant differences between this method and LDL cholesterol calculated from Friedewald's formula. CONCLUSIONS: Direct homogeneous assay for HDL cholesterol determination in diabetic patients seems not to exhibit a negative bias, in contrast to the precipitation method, when compared with the ultracentrifugation method. In addition, the direct assay saves time and is not influenced by type of diabetes or degree of metabolic control.     

两种清除法测定高密度脂蛋白胆固醇的临床评价

目的对反应促进剂过氧化物酶清除法（SPD）和过氧化氢酶清除法（CAT）两种高密度脂蛋白胆固醇（HDL—C）均相测定法进行临床评价。方法将上述两种方法与磷钨酸镁法（PTA—Mg）进行比较，分析各自方法的精密度、准确性、特异性和干扰因素。结果两种清除法与PTA—Mg具有良好的相关性，SPD y1=0．9412x＋0．1123，r=0．9812；CAT y2=0．9245x＋0．1256，r=0．9845。高、中、低3种HDL-C浓度混合血清所测定结果表明两种方法均具有良好的精密度，总CV值SPD法3．35％～3．87％，CAT法3．45％～3．96％，均达到临床满意的程度。两法线性范围均较宽（线性均至4．02mmol／L），最低检测浓度均为0．08mmol／L，平均回收率SPD法为98．1％，CAT法为97．5％。三酰甘油（TG）％15．3mmol／L，血红蛋白（Hb）〈5g／L，低密度脂蛋白胆固醇（LDL-C）〈8．0mmol／L，胆红素小于450μmol／L对两法基本无影响。结论两种HDL-C清除法测定结果的准确度和精密度均符合临床要求，适宜自动分析，值得在临床推广应用。     

美国CDC胆固醇参考方法和高密度脂蛋白胆固醇指定比对方法的运行和应用

目的 运行美国CDC胆固醇(TC)参考方法和高密度脂蛋白胆固醇(HDLC)指定比对方法,用于我国血脂分析系统标准化.方法 运行CDC TC参考方法和HDLC指定比对方法,加入CDC胆固醇参考方法实验室网络(CRMLN),每年6次参加CRMLN分析质量考核,按规定程序对我国21种TC或HDLC分析系统进行认证实验.结果 TC分析在18次考核中本室平均变异系数(CV)为0.29%,相对于CDC靶值的平均偏倚小于0.1%;HDLC分析在17次考核中本室平均标准差为0.010 mmol/L(0.39 mg/dl),相对于CDC靶值的平均偏倚-0.019 mmol/L(-0.72 mg/dl),相对于组均值的平均偏倚-0.006 mmol/L(-0.25 mg/dl);本室绝大多数(>90%)TC和HDLC分析事件符合CRMLN精密度和准确度要求;21种分析系统中的18种满足CRMLN性能要求,通过CDC溯源认证.结论 与国际参考实验室网络相联系的TC参考方法和HDLC指定比对方法以及以此为基础的血脂分析系统认证计划已经建立,可望在我国血脂分析标准化中发挥重要作用.     

化学沉淀法测定小而密低密度脂蛋白方法的建立

目的利用化学沉淀原理建立小而密低密度脂蛋白(sdLDL)快速、简便的检测方法。方法通过比较不同种类及不同浓度沉淀剂对血清样本中脂蛋白的分离效果,确定最终沉淀剂,并对化学沉淀法测定血清sdLDL-C的精密度、线性范围和回收率进行分析。结果最终选择140 U/mL肝素-90 mmol/L Mg2+作为沉淀剂测定血清sdLDL胆固醇(sdLDL-C)含量。化学沉淀法测定sdLDL-C在0.14~1.44 mmol/L范围内线性关系良好,回归方程为Y=1.025X-0.197,r=0.988;对高、低2组不同sdLDL-C浓度的样本批内变异系数(CV)分别为2.83%、3.19%,批间CV分别为5.49%、6.13%;回收率为83.81%。结论 sdLDL化学沉淀法具有样本用量少、操作简便、成本低廉等优点,更适合常规实验室检测血清sdLDL-C含量。     

A new liquid homogeneous assay for HDL cholesterol determination evaluated in seven laboratories in Europe and the United States.

We evaluated a new liquid homogeneous assay for the direct measurement of high density lipoprotein cholesterol (HDL-C Plus) in seven laboratories. The assay includes two reagents which can be readily used in most available clinical chemistry analyzers. The total CVs of the new method were below 4.6% and the bias in relation to the designated comparison method was below 3.9%. The total error ranged between 4 to 7%. HDL-C values determined by this method were in good agreement with those obtained by the old homogeneous assay using lyophilized reagents, and other homogeneous and precipitation assays (0.944 < r < 0.996). The assay was linear up to at least 3.89 mmol/l HDL-C. Hemoglobin did not interfere, whereas in icteric samples slight deviations were observed. Lipemia up to 11.3 to 22.6 mmol/l triglycerides did not interfere with this homogeneous HDL-C assay. In samples of patients with paraproteinemia, discrepant results were seen. This liquid homogeneous HDL-C assay was easy to handle and produced similar results in all laboratories participating in this study. This method will enable clinical laboratories to reliably measure HDL-C for risk assessment of coronary heart disease.     

Determination of LDL cholesterol and LDL apolipoprotein B following precipitation of VLDL in blood serum with phosphotungstic acid/MgCl2

A method is described for the selective precipitation of VLDL in blood serum using phosphotungstic acid/MgCl2. The method allows for the calculation of LDL apolipoprotein B as well as for the calculation of LDL cholesterol (following the additional determination of HDL cholesterol). Dependent on the triglyceride and the cholesterol content of the serum, three different procedures were developed using phosphotungstic acid and MgCl2 in different concentrations in the precipitation assay. Within the tested range of 3-10 mmol/l total cholesterol and 1-4 mmol/l triglyceride in blood serum the VLDL were nearly completely precipitated with negligible coprecipitation of LDL and HDL, but 40-50% coprecipitation of Lp(a). Regression analysis of the cholesterol values obtained by precipitation with phosphotungstic acid/MgCl2 (= serum cholesterol - LDL cholesterol), and the cholesterol values obtained by ultracentrifugation (d greater than 1.006 kg/l) revealed a good measure of agreement (r = 0.97, y = 0.93 X + 0.35, n = 76). An equally good measure of agreement was found for the corresponding apolipoprotein B values (r = 0.96, y = 1.03 X - 0.2, n = 61). In the determination of LDL cholesterol a variation coefficient of 4.3% (n = 20) was found in relation to the precision in the series, and a variation coefficient of 4.8% (n = 25) in relation to day to day precision.     

Homogeneous HDL-cholesterol assay versus ultracentrifugation/dextran sulfate-Mg2+ precipitation and dextran sulfate-Mg2+ precipitation in healthy population and in hemodialysis patients.

OBJECTIVES: To evaluate the analytical performance of a new homogeneous HDL-cholesterol assay (Olympus Diagnostica). To investigate possibly discrepant results in chronic hemodialysis patients who commonly exhibit quantitative and qualitative lipoprotein abnormalities, responsible for atherogenic complications in these patients. DESIGN AND METHODS: Serum samples were collected from 50 healthy subjects and 65 chronic hemodialysis patients. HDL-C levels measured by the homogeneous assay were compared with the routine dextran sulfate-Mg2+ precipitation method and the ultracentrifugation/dextran sulfate-Mg2+ precipitation as reference method. RESULTS: The homogeneous assay was linear up to at least 220 mg/dL. The analytical precision was estimated with three different sets of commercial controls and one set of human pooled serum control. The within-day CV ranged between 1.7% and 3.8% and the between-day CV ranged between 1.0% and 2.3%. HDL-C values in both populations correlated highly with the dextran sulfate-Mg2+ precipitation method and the ultracentrifugation/dextran sulfate-Mg2+ precipitation method (r > or = 0.96, bias between -0.9 and 2.3 mg/dL). Lipemia up to triglyceride concentration of 600 mg/dL did not alter the HDL-C value. CONCLUSIONS: The homogeneous assay for HDL-C (Olympus) uses much less sample, is accurate and convenient to handle, and allows full automation. The test should considerably facilitate the screening of individuals at an increased risk of cardiovascular disease, including hemodialysis patients.