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.     

Quick assay of serum HBs antibody levels using latex agglutination-integrating sphere turbidimetric assay (ISTA)

A method for determining serum HBs antibody applying the principle of integrating sphere turbidimetric assay (ISTA) by latex agglutination was developed. The minimum detectable level of HBs antibody by this method is 12.5 IU/L, indicating that this method is 3 times or more sensitive with better reproducibility and specificity than the passive hemagglutination (PHA) method. With a cut-off level of 25 IU/L, the possible highest simultaneous reading by this method was 1,000 IU/L Serum HBs antibody can be readily measured in 10 or so minutes by this method if a fully automated EL-1000 analyzer is used. This rapid and simple method for determining serum HBs antibody will be useful not only clinically, but also in preventive medicine. © 1993 Wiley-Liss, Inc.     

T细胞斑点试验联合隐球菌乳胶凝集试验对肺结核合并肺隐球菌病诊断的初步研究

目的 评价结核分枝杆菌感染T细胞斑点试验(TB ELISPOT)和乳胶凝集试验(LA)联合检测对肺结核合并肺隐球菌病的诊断价值. 方法 选择2006年3月-2008年9月我中心呼吸科收治的疑诊肺结核合并肺隐球菌病患者76例,均同时作TB ELISPOT、LA检测和肺组织病理检查.结果 76例患者中,经病原学和病理学确诊为肺隐球菌病15例,肺结核22例,其中肺结核合并肺隐球菌病8例.经检测LA的灵敏度和特异度均为100%,TB ELISPOT的灵敏度和特异度分别为91%和94.4%,8例肺结核合并肺隐球菌病患者LA和TB ELISPOT均为阳性.结论 LA和TB ELISPOT联合检测对肺结核合并肺隐球菌病具有诊断价值.     

Simultaneous automated measurement of serum total CK and CK-MM lsoform ratio in serum

We automated a two-step kinetic procedure for determining serum CK-MM isoform ratio using an immunoinhibition method. By measuring the total CK activity and the residual CK activity (serum CK-MM isoform) remaining after the inhibition by tissue CK-MM isoform specific monoclonal antibody reagent (CK-M01) the CKMM isoform ratio is calculated using the difference between total CK and residual CK activities divided by the residual CK activity. Linearities of total CK and residual CK assays were?7750 U/L and 2,500 U/L, respectively; within-run CVs of isoform ratio (N = 10) were 2.8 and 7.0% (mean 0.14 and 0.60), respectively. The MM³/MM¹ isoform ratio obtained with the proposed method (X) correlated well with the results of electrophoretic method (Y) according to the equation: Y = 0.98X ?0.3, r = 0.988. The normal reference range of isoform ratios obtained by assaying 1,222 serum samples from healthy subjects was 0.09–0.75. The isoform ratio increased after onset of chest pain, peaking at 2–6 hr thereafter. A mean isoform ratio of 1.86 was obtained with serum sample from 86 patients diagnosed as having an acute myocardial infarction (AMI). This method is accurate and highly sensitive, as the detection and early diagnosis of AMI can be completed in 10 min. © 1994 Wiley-Liss, Inc.     

乳胶快速凝集与常规培养方法检测阴道毛滴虫的比较研究

目的　用阴道毛滴虫的粘附蛋白制备单克隆抗体,并研制成乳胶凝集试剂,在临床上初步应用。方法 　从阴道中收集分泌物,分别用乳胶凝集法和常规培养方法对197份妇科门诊患者的阴道分泌物标本进行检 测。结果　与常规培养方法比较,乳胶凝集法的敏感性为89.9%,特异性为95.5%,阳性预测值为68.0%,阴性 预测值为98.8%。结论　该单克隆抗体研制成的乳胶凝集法快速、简便,具有较高的特异性和较好的敏感性,能 提高临床滴虫性阴道炎的诊断率。     

急性心肌梗死患者血清缺血修饰清蛋白和肌红蛋白检测水平分析

目的分析检测缺血修饰清蛋白(IMA)和肌红蛋白(MYO)水平在急性心肌梗死(AMI)早期诊断中的临床应用价值。方法在贝克曼库尔特AU680全自动生化分析仪上检测本院47例AMI患者和42例健康者(对照组)的血清IMA、MYO、乳酸脱氢酶(LDH)、肌酸激酶(CK)及肌酸激酶同工酶(CK-MB)的水平,同时检查心电图,并对其检测结果进行统计学分析比较。结果 AMI患者发生胸痛后0~6h,血清IMA和MYO水平明显高于对照组(P0.01),血清LDH、CK和CK-MB水平也高于对照组(P0.05);胸痛6h以前血清IMA和MYO水平明显高于胸痛6h以后(P0.05),而LDH、CK和CK-MB水平差异无统计学意义(P0.05)。实验组IMA和MYO水平明显高于对照组(P0.01),血清LDH、CK和CK-MB高于对照组(P0.05)。在AMI早期(0~6h)血清IMA和MYO阳性检出率均在78.0%以上,明显高于心电图、LDH、CK和CK-MB的检出率(P0.05)。结论 IMA和MYO可作为AMI早期诊断较理想的心肌缺血指标,且出现时间早、阳性检出率高。联合检测有助于AMI的早期确诊,为病情鉴别、治疗以及预后等提供可靠依据。     

Semi-quantitative estimation of serum myoglobin by a rapid latex agglutination method: an emergency screening test for acute myocardial infarction

The present study reports the evaluation of a new latex agglutination test for serum myoglobin (SMb). The time of agglutination of the latex particles coated with antibodies to myoglobin was measured in 172 serum specimens with known concentration of myoglobin quantitated by a radioimmunoassay (RIA), collected from myocardial infarction (MI) patients, subjects suffering from various diseases, and normal controls. Myoglobin levels in the samples were found to decrease exponentially with time of agglutination. Agglutination occurring within 1 min (result coded as + + + +) corresponded to 761 +/- 366 micrograms/l of myoglobin; between 1 and 2 min (+ + +), to 285 +/- 101 micrograms/l; between 2 and 3 min (+ +), to 85 +/- 47 micrograms/l; between 3 and 4 min (+), to 51 +/- 38 micrograms/l; and after more than 4 min (-), to 31 +/- 16 micrograms/l. Blood samples were serially drawn from 24 MI patients with short hospitalization delays; the rapid agglutination which was obtained in the specimens taken upon admission (20 results coded as + + + + and four as + + +) actually corresponded to markedly increased SMb levels. In contrast, serum creatine kinase (CK) activities were still less than 150 U/l in four patients (16.6%); CK-MB was less than 5 U/l in five cases (20.8%). Positive agglutinations for SMb were also obtained 4 and 8 h following admission in all subjects, confirming that the latex test is an early and very sensitive indicator for MI.(ABSTRACT TRUNCATED AT 250 WORDS)     

A latex agglutination test for rapid quantitative estimation of the plasmin–antiplasmin complex in human plasma

An antiserum was raised in rabbits against human plasmin-antiplasmin complex and rendered specific for neoantigens of this complex by absorption with purified plasminogen and plasma. Polystyrene particles were coated with the specific antibodies and used in an agglutination test for the determination of plasmin-antiplasmin complex in the plasma from various patients. Purified plasmin-antiplasma complex at a concentration of 0.1-0.2 mg/l was found to cause a clear agglutination of the particles. Activation of fresh human plasma with urokinase caused progressive generation of agglutinating activity up to a plasma dilution of 1/480. Intravenous infusion of streptokinase into patients resulted in an increase of the plasmin-antiplasmin titre of at least 1/240. Sera from patients with rheumatoid factor also agglutinated the particles but this activity could be removed by absorbing rheumatoid factor on insolubilized human IgG. Out of 101 male and twenty-three female control subjects, only three men had a plasmin-antiplasmin titre above 1/16. Of 230 hospitalized patients, plasmin-antiplasmin titres of 1/40 or more were detected in twenty-five patients. Most of these patients had diseases which are frequently associated with in vivo coagulation or fibrinolysis, but among them there was only one who showed diffuse intravascular coagulation detectable by classical methods. In the absence of an increased plasmin-antiplasmin titre none of the haemostasis analyses were indicative of in vivo coagulation or fibrinolysis. Seven out of eight patients with diffuse intravascular coagulation of various origin had plasmin-antiplasmin titres of 1/80 or 1/160. Thus, the present latex agglutination test, owing to its simplicity and sensitivity, appears to be a practical routine screening test for detecting fibrinolytic activation in plasma.     

Quantitative automated latex nephelometric immunoassay for determination of myoglobin in human serum

We have evaluated a new latex nephelometric test for the quantitation of myoglobin in human serum. The assay consists of incubating the diluted serum sample (20-fold) for 12 min at room temperature with latex particles covalently coated with anti-myoglobin antibodies and then quantifying the change of light-scatter produced. The assay is fully automated on the Behring nephelometer analyzer with a sampling rate of 150 samples/hour. There is no interference from bilirubin (up to 340 mumol/l), haemoglobin (up to 7,000 mg/l), or rheumatoid factor (up to 1,100 int. units/ml). Myoglobin standard curve extends from 20 to 380 micrograms/l. Assay detection limit lies around 6 micrograms/l. Coefficient of variation ranged from 2.7 to 7.6%. Correlation coefficient between latex immunoassay and an RIA method was 0.987, calculated from the assay of 37 samples. A statistically significant difference was found between the distribution for females and males. The serum level of myoglobin showed an age-dependent variation. Concentrations up to 60 micrograms/l are considered to be normal.     

A new immunoassay for the measurement of myoglobin in serum

Because the concentration of serum myoglobin (Mb) increases within 2 to 4 hours after the first sign of acute myocardial infarction, it has been proposed as an early marker of the condition. Our aim was to evaluate a new assay that provides a rapid, quantitative determination of Mb (Baxter Stratus Myoglobin) based on the radial partition technique. We compared the results obtained by this technique with those from nephelometric and radioimmunoassay methods. A significant agreement was observed, the correlation coefficients (r) being 0.999 and 0.996, respectively. The method evaluated provided good reproducibility with CVs between 3.14% and 4.87%, and its linearity and analytical sensitivity were satisfactory. The clinical evaluation of this assay demonstrates that Mb increases in serum of patients with acute myocardial infarction before total creatine kinase and creatine kinase MB isoenzyme. Mb concentration shows an early peak and earlier return to normal values after the necrosis compared to enzymatic activities. Moreover the assay is rapid and fully automated. The method is therefore considered appropriate for contributing to the early diagnosis of AMI in clinical laboratories. © 1994 Wiley-Liss, Inc.     

一种新的纤溶活性测定法及在心肌梗死中的应用

目的 建立新的纤维蛋白溶解活性（Ｆｉｂｒｉｎｏｌｙｔｉｃａｃｔｉｖｉｔｙ,ＦＬＡ）测定方法,并初步应用于心肌梗死（ＡＭＩ）患者的观察,方法 将定量受检血浆加于含有纤溶酶原（ＰＬＧ）和标准纤维蛋白的小试管中,同时加入定量的组织纤溶酶原激活剂（ｔ－ＰＡ）硅粒,３７℃温育一小时,检测生成Ｄ－二聚体的量,结果 ４０例正常人的ＦＬＡ分级在Ⅱ,Ⅲ级（占７０％）,６０例ＡＭＩ患者ＦＬＡ分级在０,Ｉ和Ⅳ级（占７３     

Development of a rapid microparticle-enhanced nephelometric immunoassay for serum myoglobin in acute myocardial infarction.

A microparticle-enhanced nephelometric immunoassay was developed for myoglobin quantitation in human serum. It uses rabbit antimyoglobin serum and hydrophilic polyacrylic microparticles covalently coated with baboon myoglobin in a competitive immunoagglutination system. The level of microparticle agglutination is assessed with a specially designed nephelometer. This sensitive (45 ng/ml of myoglobin detected in serum) and accurate (coefficients of variation from 3.0% to 8.2% in precision study and linear recovery of myoglobin in overloaded sera) immunoassay was evaluated with human sera from patients suffering from acute myocardial infarction. Myoglobin levels in patient's serum on admission appeared to be correlated with clinical and biological parameters assessed in emergency wards and later. This new, rapid, and easy microparticle-enhanced nephelometric immunoassay could thus be useful in emergency conditions for the early quantitation of serum myoglobin.     

Automated quantitative nephelometric latex immunoassay for determining ferritin in human serum.

We describe a rapid and sensitive latex nephelometric immunoassay for quantifying ferritin in human serum. This latex immunoassay procedure uses commercially available ready-for-use reagents [Tina-Quant (a) Ferritin, Boehringer Mannheim] that have a long shelf life. The assay consists of incubating the diluted serum sample (5-fold) for 12 min at room temperature with latex particles covalently coated with anti-ferritin antibodies, and then quantifying the change of light-scatter produced. The assay is fully automated on the Behring nephelometer analyzer with a sampling rate of 150 samples/hour. The method has an analytical range of 3 to 260 micrograms/l. Maximal intra- and inter-assay CVs were 4.0 and 6.2%, respectively. Analytical recoveries ranged from 91.3 to 103.6%. Assay detection limit was less than 3 micrograms/l. Linearity of the test is given throughout the measuring range. There was no interference from bilirubin (up to 340 mumol/l), haemoglobin (up to 7 g/l), or rheumatoid factor (up to 1,100 IU/ml). Turbid and lipemic samples interfere. This interference may be avoided by pretreating these samples prior to assay. Results correlated well with those obtained by an automated ELISA test (r = 0.995) and with those of two commercial RIA methods (r greater than 0.97). This latex nephelometric procedure is a convenient method and represents an interesting alternative to other immunoassays for measuring ferritin in human serum.     

Automated measurement of a constitutive isotype of serum amyloid A/SAA4 and comparison with other apolipoproteins

A constitutive isotype of serum amyloid A (SAA4) is present mostly in high density lipoprotein (HDL) and a little in other lipoproteins. In this study, we developed an automated method for measuring SAA4 concentration in serum by kinetic nephelometry of anti-SAA4 antibody-coated latex agglutination. Rabbit antibodies generated by immunization of recombinant SAA4 were found to have no apparent reactivity with acute phase SAA. The values determined by this method and by the previous enzyme immunoassay showed good agreement (r=0.862). Serum SAA4 values of 26 healthy adults ranged from 37–109 mg/L (mean; 62 mg/L). Their SAA4 concentrations were not significantly related with those of apolipoprotein A-I, A-II, B, C-II, C-III or E. Also, SAA4 did not correlate with cholesterol in preparation after removal of very low density lipoprotein and low density lipoprotein. These suggest the unique behavior of SAA4 in lipoprotein metabolism, while what contributes to variation of SAA4 levels in serum, especially in HDL, remains to be clarified. J. Clin. Lab. Anal. 11:363–368, 1997. © 1997 Wiley-Liss, Inc.     

Measurement of antigen concentration by an ultrasound-enhanced latex immunoagglutination assay

A novel ultrasonic technique that increases the rate and sensitivity of latex agglutination tests (LATs) has recently been described. The technique is based on the fact that suspended latex particles become concentrated in an ultrasonic standing wavefield, thereby increasing the rate of particle-particle collisions compared to the standard LAT procedure. The present work extends earlier qualitative assessments of agglutination and seeks to establish whether quantitative measurement of agglutinate size may be used as an indicator of antigen concentration. The agglutination of latex microparticles coated with antibody to C-reactive protein (CRP) is studied here as a model system to determine the dependence of agglutinate size on analyte (CRP) concentration. Agglutinate size is characterised by image analysis techniques. The results show that agglutinate size decreases with decreasing CRP concentration. A near linear relationship is shown between analyte concentration and the size of agglutinate formed over a 100-fold dilution range. The threshold concentration of 230 pg/mL for detection of CRP in the ultrasonic test is 2560 times lower than that required for a conventional test-card CRP latex agglutination assay.     

联合检测肌钙蛋白T及肌红蛋白和NT-ProBNP在心肌梗死诊断中的临床研究

目的观察联合检测肌钙蛋白(cTnT)、肌红蛋白(Myo)及血浆氨基末端B型利钠肽前体(NT-proBNP)在心肌梗死(MI)诊断中的临床应用。方法选择2012年1月至2015年12月以急性胸痛为主诉入院进行检查治疗的患者共计913例,根据最终诊断结果将患者分为急性MI组和非急性MI组。所有患者来诊后检测其cTnT、Myo及NT-proBNP水平。观察2组患者cTnT、Myo、NT-proBNP的差异,观察上述3项指标单独检测、二联检测和三联检测诊断急性MI的灵敏度和特异度。结果急性MI组患者cTnT、Myo、NT-proBNP水平均明显高于非急性MI组,差异有统计学意义(P0.05)。二联检验的灵敏度和特异度高于cTnT、Myo、NT-proBNP单独检验,而三联检验的灵敏度为95.18%,特异度为89.86%,明显高于单独检验及二联检验。结论 cTnT、Myo、NT-proBNP的联合检验在MI诊断中具有重要意义,能够提高MI诊断的特异度和灵敏度,为临床治疗提供可靠依据。