免疫透射比浊法测定血清轻链

目的：评价采用免疫透射比浊法检测血清轻链的可行性。方法：在贝克曼CX4全自动生化分析仪上,应用ROCHE公司试剂,以免疫透射比浊法,采用二点终点法,主波长340nm,副波长700nm,温度37℃,六点校准。结果：本法检测kappa轻链,日内不精密度2.5～2.8,日间不精密度3.0～3.2,检测低限为0.19g/L,回收率为92.1～101.5,检测lambda轻链,日内不精密度1.6～1.8,日间不精密度2.2～2.5,检测低限为0.16g/L,回收率为94.2～102.5,免疫透射比浊法（Y）与贝克曼IMAGE特定蛋白分析仪的免疫散射比浊法（X）比较：kappa轻链Y=0.265X-0.232。决定系数（r^2）=0.957,lambda轻链Y=0.438X-0.03。决定系数（r^2）=0.976。结论：免疫透射比浊法简便,灵敏,试剂稳定,适合一般医院血清轻链的自动化分析。     

免疫透射比浊法测定尿轻链

免疫球蛋白（Ig）是血清中重要的蛋白质,尿液中也有微量存在。免疫球蛋白由二条重链和二条轻链组成,重链有γ、α、μ、δ、ε五种,分别对应于IgG、IgA、IgM、IgD、IgE,轻链有二种,每个免疫球蛋白分子或含kappa轻链,     

免疫透射比浊法测定血清IgE的方法建立与评价

目的：评价采用免疫透射比浊法检测血清免疫球蛋白E（IgE）的可行性。方法：在贝克曼CX4全自动生化分析仪上,应用德赛公司试剂,以免疫透射比浊法,采用二点终点法,主波长560nm,副波长700nm,温度37℃,六点校准。结果：本方法线性范围（50～960）IU/ml,日内不精密度1.02%～2.56%,日间不精密度1.24%～5.61%,检测低限为12.7IU/ml,回收率为93.2%～102.5%,免疫透射比浊法（Y）与贝克曼IMAGE特定蛋白分析仪的免疫散射比浊法（X）比较：Y=0.959X＋4.623。决定系数（r2）=0.9883。结论：免疫透射比浊法简便、灵敏、试剂稳定,适合一般医院血清免疫球蛋白E的自动化分析。     

免疫透射比浊法测定肿瘤特异性生长因子方法学评价

对肿瘤特异性生长因子（TSGF）方法学和临床应用进行评价,采用免疫透射比浊法与微粒子酶免疫法（MEIA）进行比较,对其重复性、线性等进行分析。用本法与MEIA法检测120份血清标本TSGF浓度,结果经χ^2检验,无显著性差异（P〉0．05）,与MEIA法比较阳性符合率为95．7％,阴性符合率为96％,总符合率为95．8％;低、高浓度的批内CV为3．8％和4．2％,批间CV为4．0％和4．9％;检测浓度小于64U／mL时,线性良好,Y=0．899X＋1．876,r=0．9882;RF、SLE和炎症患者血清和重度溶血、脂血、黄疸血清等对检测均有～定的影响。该TSGF方法符合临床常规实验室检测的要求。     

化学发光法与免疫透射比浊法检测血清纤维连接蛋白的方法评价

目的分析化学发光法与免疫透射比浊法测定血清纤维连接蛋白（FN）的偏倚,为临床实验室选择项目提供一定的依据。方法根据美国临床实验室标准化研究所（CLSI）EP9-A2文件提供的实验流程,每天随机选取临床标本8份,分别用两种方法进行检测,共测定6d,记录实验结果。结果以免疫透射比浊法（X）为参比方法,对化学发光法（Y）进行评估。两种方法测定的回归方程式为y=1.034x-1.308,r2=0.958。当免疫透射比浊法的浓度为100、200、300mg/L时,它们的相对偏倚为2.1%、2.7%、3.1%。结论化学发光法与免疫透射比浊法测定偏倚比较评价均可接受,两种方法检测血清FN均具有良好的相关性,建议各临床实验室对不同方法建立参考值范围。     

免疫透射比浊法血清白蛋白检测试剂盒的性能验证和临床应用评价

目的对免疫透射比浊法血清白蛋白检测试剂盒进行性能验证和临床应用评价。方法根据美国临床实验室标准化委员会(CLSI)的标准要求,使用全自动日立生化分析仪7600-020,对该试剂盒的精密度、正确度、检出限、线性范围、干扰实验、参考区间、方法学比对进行验证,并将检测结果和厂家提供的的性能指标进行比较。结果该试剂检测血清白蛋白的批内精密度为1.1%、批间精密度为1.2%,均小于厂家提供的指标;正确度在允许偏倚范围内;检出限为0.152g/L;线性斜率为0.9755,相关系数的平方R2为0.9945,大于0.99符合要求;生物参考区间符合率100%;在厂家说明书标示的干扰物浓度范围内,对检测结果没有明显影响;与溴甲酚绿法做比对实验,相关性回归方程为Y=0.9894X-1.1033,R2为0.9794,在低值结果中两种方法学差异呈非线性分布,无法用校正系数调整。结论免疫透射比浊法血清白蛋白检测试剂盒在7600-020生化分析仪上主要分析性能符合厂家的声明,结果准确可靠,抗干扰能力强,可以开展用于临床检验分析。     

胶乳增强免疫透射比浊法测定CRP的方法学评价

CRP的测定方法主要有免疫比浊法、免疫单抗法、胶乳凝集法、火箭电泳法,其中以免疫特定蛋白仪的免疫散射比浊法作为参考方法,而其他几种方法操作繁琐又不易于自动化分析。胶乳增强免疫透射比浊法是近年来发展较为可靠的方法,具有检测敏感度高、线性范围宽、结果稳定、     

免疫透射比浊法检测尿微量白蛋白的方法学评价

近来年研究表明,尿微量白蛋白不仅是肾脏血管内皮损伤的标志,也是全身血管内皮细胞损伤的标志。一些调查研究认为,尿微量白蛋白阳性是心血管事件发生的独立危险因素[1]。因此,检测尿微量白蛋白对相关疾病的早期诊断和治疗有重要的参考意义。为了验证免疫透射比浊法（简称＂透射法＂）微量白蛋白诊断试剂用于尿微量白蛋白检测的准确性及实用性,本文以免疫散射比浊法（简称＂散射法＂）微量白蛋白诊断试剂作参比方法,对检测结果进行相关性分析和方法学评价。     

颗粒增强免疫透射比浊测定血中Cyst C全自动分析法的建立与评价

目的：建立颗粒增强免疫透射比浊测定血中胱蛋白酶抑制剂C（Cyst C）全自动分析,评价该法常规用于检测血中胱蛋白酶抑制剂C的可行性。方法：将含有Cyst C血样本与乳胶颗粒增强的Cyst C多克隆兔抗体按一定体积比混合,在一定量的兔免疫球蛋白及表面活性剂存在条件下,抗原、抗体结合反应在一定温度、时间内产生一定大小的颗粒,许多颗粒在反应液体中形成一定的浊度,利用Olympus AU2700全自动生化分析仪透射比浊测定,并对方法的不精密度、准确度、干扰试验及与Dade Behring BN ProSpeC免疫散射比浊测定方法比较等进行评价。结果：Cyst C浓度为1．03mg／L、5．67mg／L样本的批内不精密度CV分别为4．8％、1．9％。向Cyst C浓度为0．79mg／L、1．18mg／L、1．46mg／L、2．46mg／L样本中加入1．65mg／L的标准液测定回收率分别为100％、95％、85％、87％,平均回收率为92％。样本中的类风湿因子、胆红素、血红蛋白、三酰甘油等干扰物浓度分别达975U／ml、400μmol／L、5g／L、6．7mmol／L时对检测无显著性影响。Cyst C浓度在0．41mg／L～6．60mg／L范围内检测线性良好。血清、肝素抗凝血浆及EDTA抗凝血浆样本测得的Cyst C结果未发现显著性差异（F=0．065,P=0．938）。与Dade Behring BN ProSpeC免疫散射比浊测定方法比较有良好的相关性,Y=0．98x-0．054,r=0．997,P〈0．01,但两者存在非常显著性差异（P〈0．01）,本法结果较Dade Behring BN ProSpeC免疫散射仪测定结果略低,两种方法的平均偏差为-0．0678mg／L。结论：建立的颗粒增强免疫透射比浊法测定血Cyst C重复性好、准确度高。血清、肝素抗凝血浆及EDTA抗凝血浆均可用于分析。测定不受类风湿因子（≥975U／ml）,胆红素（≥400μmol／L）,血红蛋白（≥5g／L）及三酰甘油（≥6．7mmol／L）的干扰。与Dade Behring BN ProSpec免疫散射比浊测定方法测定结果比较有良好的相关性,适用于在全自动生化分析仪上作常规检测。     

免疫透射比浊法测定血清甘胆酸

本文在全自动生化分析仪上,以免疫透射比浊法测定甘胆酸(cholyglycine,CG),取得满意结果,特作报道.1 材料和方法1.1 材料1.1.1 样本 来源于浙江大学医学院附属第一医院临床血清标本(来源于肝炎病房和肝胆胰外科中心),肝癌24例,慢性肝炎41例,急性肝炎12例,肝硬化21例,梗阻性黄疸7例,合计105份标本.35例正常人样本来自本院体验标本(排除肝病且肝功能全部正常的标本).     

Hypocretin-1 measurements in cerebrospinal fluid using radioimmunoassay: within and between assay reliability and limit of quantification

Study ObjectivesThe most sensitive and specific investigative method for the diagnosis of narcolepsy type 1 (NT1) is the determination of hypocretin-1 (orexin-A) deficiency (≤110 pg/mL) in cerebrospinal fluid using a radioimmunoassay (RIA). We aimed to assess the reliability of the Phoenix Pharmaceuticals hypocretin-1 RIA, by determining the lower limit of quantification (LLOQ), the variability around the cutoff of 110 pg/mL, and the inter- and intra-assay variability.MethodsRaw data of 80 consecutive hypocretin-1 RIAs were used to estimate the intra- and inter-assay coefficient of variation (CV). The LLOQ was established and defined as the lowest converted concentration with a CV <25%; the conversion is performed using a harmonization sample which is internationally used to minimize variation between RIAs.ResultsThe mean intra-assay CV was 4.7%, while the unconverted inter-assay CV was 28.3% (18.5% excluding 2 outliers) and 7.5% when converted to international values. The LLOQ was determined as 27.9 pg/mL. The intra-assay CV of RIAs with lower specific radioactive activity showed a median of 5.6% (n = 41, range 1.6%–17.0%), which was significantly higher than in RIAs with higher specific activity (n = 36; median 3.2%, range 0.4%–11.6%, p = .013). The CV around the 110 pg/mL cutoff was <7%.ConclusionsHypocretin-1 RIAs should always be harmonized using standard reference material. The specific activity of an RIA has a significant impact on its reliability, because of the decay of ¹²⁵I radioactivity. Values around the hypocretin-1 cut-off can reliably be measured. Hypocretin-1 concentrations below 28 pg/mL should be reported as “undetectable” when measured with the Phoenix Pharmaceuticals RIA.Clinical Trial InformationThis study is not registered in a clinical trial register, as it has a retrospective database design     

Analytic accuracy of specific immunoglobulin E antibody results determined by a blind proficiency survey

Analytic variability affects the accuracy of measurements of specific IgE antibodies, but the frequency of false results attributable to analytic variability is not well documented. We have monitored the accuracy of the results generated in our laboratory by testing aliquots of positive serum pools and a negative serum pool submitted blindly for the measurement of IgE antibodies to 16 different allergens, including foods; weed, grass, and tree pollens; mites, molds, and epithelia; and Hymenoptera venoms. Positive serum pools were prepared to contain modest amounts of IgE antibodies. Tests were performed by immunometric assays with microcrystalline cellulose allergen immunosorbents. Frank false-positive and false-negative results were very uncommon when binding levels were classified by a ratio-based reporting scheme. False-borderline results were more common. A borderline result is truly equivocal and may or may not indicate the presence of low levels of IgE antibodies. Analytic variability adds uncertainty to the measurement of small quantities of IgE antibodies regardless of the classification scheme used to report test results.     

Design of a portable urine glucose monitoring system for health care

This paper describes the design of a monitoring system that can be used to measure urine glucose during daily life. It consists of a bio-chemical sensor, hardware with PIC microcontroller and control circuits, and signal analyzing part. To evaluate the performance, we compared the analyzed glucose levels of the developed system to a standard instrument, YSI glucose analyzer, based on regression analysis using standard glucose solutions mixed with urine. Also, standard deviation and coefficient of variation were computed. In conclusion, the developed system showed it could be used for the measurement of urine glucose.     

Clinical significance of serum LpA-I levels measured by immunoturbidimetric assay in type 2 diabetes mellitus patients

Previously, we developed an immunoturbidimetric assay method for lipoprotein A-I(LpA-I) on sera pre-absorbed with anti-apolipoprotein A-II. In the present study, correlations between serum lipoprotein A-I and other serum parameters levels were examined and LpA-I levels were studied in patients with type 2 diabetes mellitus. The serum levels of LpA-I did not correlate with those of diabetic markers such as fasted blood glucose, glycohemoglobin(HbA1c) and fructosamine, but correlated well with the levels of total cholesterol and HDL cholesterol, phospholipids, apolipoprotein A-I and seemed to correlate inversely with arteriosclerosis index. In patients with type 2 diabetes mellitus, LpA-I levels were significantly lower than those in normal subjects. Especially, LpA-I levels of patients with diabetic complications were significantly lower than those in normal subjects and non-complicated diabetic patients. Then, the measurement of LpA-I levels in patients with type 2 diabetes mellitus was considered to be useful for prevention and management of arteriosclerosis.     

新鲜家兔血清对家兔急性实验性血清病的治疗作用

根据补体具有溶解免疫复合物(CMSC)作用,本文应用家兔急性实验性血清病(AESSR)动物模型,用新鲜家兔血清进行治疗。结果表明:新鲜血清可明显提高 CMSC 及 CH_(50)水平,明显降低 CIC,与阳性对照组比 P<0.01;肾脏损伤明显减轻,与阳性对照组比 P<0.05。在高峰期,大剂量组与小剂量组各项指标除 CMSC 有显著差异(P<0.01)外,其余名项均无显著差异。本文证明应用新鲜家兔血清,可明显降低 AESSR 血清中 CIC 水平,减轻家兔肾脏病理损伤,为临床应用新鲜血清治疗免疫复合物病,提供了实验基础。     

The role of serum proteins in Staphylococcus aureus adhesion to ethylene glycol coated surfaces

Bacterial adhesion on implants is a first step in the development of chronic foreign body associated infections. Finding strategies to minimize bacterial adhesion may contribute to minimize such infections. It is known that surfaces with oligo-ethylene-glycol (EG₃OMe) or poly-ethylene-glycol (PEG2k) terminations decrease unspecific protein adsorption and bacterial adhesion. However, little is known about the influence of serum and its components on bacterial adhesion. We therefore prepared two coatings on gold surface with HS-(CH₂)₁₁EG₃OMe (EG₃OMe) and PEG2k-thiol and studied the role of bovine serum albumin (BSA), γ-globulins, and serum on Staphylococcus aureus adhesion. While BSA and lysozyme showed no adherence even when applied at very high concentrations (100 mg/ml), γ-globulins adsorbed already from 10 mg/ml on. The adsorption of γ-globulins was, however, significantly decreased when it was mixed with BSA in a ratio of 3:1, as it is in the serum. Pretreatment of EG₃OMe and PEG2k coatings with γ-globulins or serum strongly promoted adherence of S. aureus when resuspended in buffer, suggesting that γ-globulins play a pivotal role in promoting S. aureus adhesion by its IgG binding proteins; the finding that a spa-deletion mutant, lacking the IgG binding protein A, showed decreased adherence corroborated this. Similarly, when S. aureus was pretreated with serum or γ-globulins its adherence was also significantly decreased. Our findings show that particularly γ-globulins bind to the coated surfaces thus mediating adherence of S. aureus via its protein A. As pretreatment of S. aureus with serum or γ-globulins significantly decreased adherence, treatment of patients with γ-globulins before implant surgery might lower the risk of implant-associated infections.     

Indirect immunofluorescence assay for the detection of hepatitis A virus-specific serum immunoglobulins.

Hepatitis A virus-specific BSC-1 cells were used for the detection of serum immunoglobulins to hepatitis A virus by indirect immunofluorescence. Of 150 serum samples tested, specific immunoglobulin M was detected only in patients with serologically confirmed acute hepatitis A, while specific immunoglobulin G was detected in patients with acute or past clinical hepatitis A as well as many patients with no known history of hepatitis.     

Localization of serum immunoglobulins in tissue of rat autonomic ganglia

The distribution of immunoglobulins is studied on preparations of autonomic ganglia using an immunohistochemical technique. Perikarya of neurons are found to be highly accessible to serum macromolecules in tissue of the vagal caudal ganglion, while myenteric ganglion tissue is devoid of immunoglobulins. An individual pattern of serum immunoglobulin distribution is typical for other ganglia. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 8, pp. 192–194, August, 1995 Presented by V. V. Kupriyanov, Member of the Russian Academy of Medical Sciences     

Immunoturbidimetric assay for estimating free light chains of immunoglobulins in urine and serum.

An immunoturbidimetric assay for the assessment of free kappa and lambda light chains of immunoglobulins was developed using a commercial polyclonal antiserum with reactivity towards epitopes on the light chains, which are not expressed when they are bound to heavy chains. The assay, on a centrifugal analyser, is simple and rapid. The limit of detection is 5 mg/l of free light chain, with an assay range of 5-120 mg/l, intrabatch precisions from 1.5-6.4%, and interbatch precisions from 6.5-8.9%. The assay was only slightly less sensitive than colloidal gold staining of cellulose acetate electrophoreses for the detection of Bence-Jones protein in urine. For the serial monitoring of response to chemotherapy in patients with myeloma, the assay correlated well with serum paraprotein estimates obtained by densitometric scanning of Ponceau stained cellulose acetate electrophoreses, but not with serum beta-2 microglobulin measurements, even after correction for the effects of creatinine. These assays may prove to be of use for the monitoring of tumour response in the treatment of Bence-Jones myeloma.     

The half-lives of serum immunoglobulins in adult mice

We determined the half-lives of several sets of murine monoclonal antibodies spanning all immunoglobulin isotypes in the serum. The antibodies in each set possess the same V region. With this approach, the differences in half-life observed between the different isotypes are independent of the V region carried by the monoclonal antibodies and therefore must relate to each other in the same way as the half-lives of each class of serum immunoglobulins. The half-life of a monoclonal antibody of the gamma 2a isotype is identical to the average half-life of serum IgG2a as previously determined (6-8 days; P. Vieira and K. Rajewsky, Eur. J. Immunol. 1986. 16:871). Therefore, the half-lives determined with monoclonal antibodies possessing the same V region represent the half-life of the serum immunoglobulins. In this way we calculated the half-life of IgM as 2 days, IgG3 and IgG1 as 6-8 days, IgG2b has a half-life of 4-6 days. IgE has a half-life of 12 h. A polymeric form of IgA was found to be eliminated from the serum with a half-life of 17-22 h.