免疫抑制法测定血清淀粉酶同工酶

本文报道了从人唾液中纯化唾液淀粉酶并制备抗血清,再利用PAMY-sepharose4B亲和层析柱从抗血清中制得特异抗SAMY抗体,进而建立了免疫抑制法来测定血清淀粉酶同工酶,并对此法的可靠性等作了探讨,并建立了成人血清淀粉酶同工酶的正常参考值。     

选择抑制法测定淀粉酶同工酶

本文对选择抑制法测定淀粉酶同工酶的实验条件作了探讨，通过数学推导和实验，建立了结果计算公式和标准曲线。本法线性范围：０－１５００ｕ／Ｌ，回收率，血清９８－１０２，尿液９２－９５，批内，批弹ＣＶ均〈９％，灵敏度：０．００１ＡＵ＝５ＩＵ／Ｌ，脂血，胆红素，葡萄糖对本实验无干扰。     

选择抑制法测定淀粉酶同工酶

在以PNPG7底物酶偶联速率法测定淀粉酶的基础上 ,对选择抑制法测定淀粉酶同工酶的实验条件作了探讨 ,通过数学推讨和实验 ,建立了结果计算公式和标准曲线。本法线性范围为 0～ 15 0 0u/L ,批内、批间CV均 <9% ,参考值范围 :血清胰型淀粉酶 (P Amy) 5 .5～ 6 7U/L ,唾液型淀粉酶 (S Amy) 1.2～ 78.2U/L ;尿P Amy4 .3～ 2 7.6U/L ,S Amy1.9～ 2 9.2U/L。本法试剂稳定期长 ,保存使用方便。选择抑制法测定淀粉酶同工酶@王建国$四川省林业中心医院检验科…     

淀粉酶同工酶琼脂糖电泳测定法

淀粉酶同工酶琼脂糖电泳测定法罗玲，赵月霞，杨振华，李云霞，贾天军（北京医院检验科，北京１００７３０）α－淀粉酶（ＥＣ．３．２．１．１，Ａｍｙ）的测定一直用于诊断胰腺疾病。但α－Ａｍｙ除来自胰腺外．还来自唾液腺，即胰型锭粉酶（Ｐ－Ａｍｙ），唾液型淀粉酶...     

淀粉酶同工酶的测定方法和临床意义

淀粉酶同工酶的测定方法和临床意义周筱云（湖北省罗田县人民医院，４３６６００）自１９６４年Ｎｏｒｂｙ从人体内α－淀粉酶中分离出同工酶以来，随着科学技术的进步，淀粉酶同工酶的测定方法越来越多，临床应用也越来越广泛，本文对这方面的进展作简要综述。１淀粉酶同...     

自制选择性淀粉酶抑制物及在淀粉酶同工酶测定中的应用

本文报道了从市售面粉中提取淀粉酶抑制物，此抑制物能比较特异地抑制唾液淀粉酶，进而建立了选择抑制法来测定血清ａ－淀粉酶同工酶，并对此法的可靠性等作了研究。建立了成人血清淀粉酶同工酶的参考值范围，并作初步临床应用。     

选择性抑制法测定AST线粒体同工酶的试验探讨

目的探讨AST线粒体同工酶(m-AST)测定的方法学评价.方法采用选择性抑制法使s-AST形成免疫复合物后失去活性,然后测出余下的m-AST.结果该法线性可达到200U/L,重复性试验批内CV＜6.5%,批间CV＜9.0%,回收率98.2%～104.4%,平均为101.4%;血红蛋白≤4 g/L,胆红素≤340 μmol/L,甘油三酯≤5.5 mmol/L和抗坏血酸≤6 g/L时对m-AST的活性测定不产生干扰作用;初步临床观察,其诊断心肌梗死的敏感性优于AST.结论m-AST测定的方法简便易行、结果准确可靠,该项目的开展对临床诊断起到了很大的促进作用.     

α—淀粉酶同工酶两种测定方法的比较

本文在建立了免疫抑制法和选择抑制法测定淀粉酶同工酶的基础上，对两法的抑制时间、抑制率、交叉抑制及正常参考值进行了比较，并认为两方法极为相关，回归极显著。     

Determinations of amylase isoenzymes in serum by use of a selective inhibitor

A simple, rapid screening procedure for determining the relative amounts of pancreatic (P)- and salivary (S)-type amylase in serum is presented, involving incorporation of a selective inhibitor (from wheat-germ) into commercially available BMD Single-Vial Amylase and Beckman Enzymatic Amylase-DS procedures for manual and automated isoamylase measurements. Optimal concentrations of inhibitor inhibit the S-type amylase fraction by 87-88%. In contrast, the pancreatic fraction is inhibited by only 19% in either the manual or automated methods. The degree of inhibition is constant for amylase activities up to at least 520 U/L. Use of the ratio (P-amylase/total amylase activity) X 100 helps differentiate between hyperamylasemia caused by S-type or P-type amylase. In preliminary studies, patients with pancreatitis showed a ratio greater than 70%.     

Automated measurement of amylase isoenzymes with 4-nitrophenyl-maltoheptaoside as substrate and use of a selective amylase inhibitor

We automated a kinetic procedure for determining amylase isoenzymes in serum and urine samples. We used 4-nitro-phenylmaltoheptaoside as substrate and a selective amylase inhibitor with the Abbott-VP bichromatic system. By use of the maximum differences between pancreatic (P) and salivary (S) amylase activities remaining after inhibition by the selective inhibitor and by use of the linear range, a one-point standard method for calibration is proposed for determining amylase activities between about 50 and 1500 U/L when the P/S ratio exceeds 0.2. Results correlated well with those by electrophoresis and the Phadebas method (r = 0.99 for both pancreatic and salivary amylase). Reproducibilities (CVs) were 1.5% to 5.5% for pancreatic amylase and 1.4% to 3.3% for salivary amylase in serum, 0.8% to 2.0% for pancreatic amylase and 0.8% to 2.3% for salivary amylase in urine.     

Effect of substrate size on immunoinhibition of amylase activity

Immunoinhibition assays are hypothesized to work by antibodies blocking substrate access to enzyme active sites. To test this hypothesis, the inhibition of amylase isoenzymes by monoclonal and polyclonal antisera was assessed using substrates of varying sizes: chromogenic sustrates 3, 5, or 7 glucose units in length, novel synthetic macromolecular substrates, and starch. The synthetic macromolecular substrates consisted of small oligosaccharide substrates linked to an inert polymer that conferred a large size to substrate molecules as determined by gel filtration chromatography. When substrate size increased, amylase activity could be inhibited equivalently by antibody concentrations that are 10-fold lower. Progressively less polyclonal serum was required to inhibit amylase activity as substrate length increased from 3 to 5 to 7 glucose units and as size was increased by linkage to a polymer. Different effects of substrate size were observed with two monoclonal antibodies. One monoclonal antibody blocked amylase activity independent of substrate size, while another monoclonal antibody had little inhibitory effect except using starch as substrate. We conclude that use of larger substrates can expand the repertoire of inhibitory epitopes on enzymes and convert a noninhibitory antibody into an inhibitory one.     

3种淀粉酶测定方法的应用评价

目的对常用的几种淀粉酶（AMY）测定方法进行临床应用评价。方法依照美国实验室标准化委员会（CLSI）EP7-A2、EP9-A2文件对3种方法（EPS法、CNPG3法和麦芽四糖法即贝克曼专用试剂）的精密度、比对试验和偏倚评估、线性分析及干扰试验进行测试。结果 3种淀粉酶测定方法的日内精密度小于1/4总分析误差,日间精密度小于1/3总分析误差。以EPS法为比较方法（X）,CNPG3法与贝克曼专用试剂为实验方法（Y）,用临床标本作比对分析,相关系数（r）分别为0.999和0.998,贝克曼专用试剂在3个医学决定水平（XC=50、120、200）处相对偏差分别为28.42%、18.35%、15.48%。CNPG3法对高脂血的抗干扰能力较EPS法与贝克曼专用试剂要高,但对混有较多红细胞的样品会出现假性增高;EPS法的线性较CNPG3法和贝克曼专用试剂要高。结论 3种方法的相关性较好,贝克曼专用试剂存在偏差,可进行一定修正。3种方法的线性与抗高脂血的干扰都能满足临床要求,CNPG3法容易受到红细胞的干扰而使结果假性增高造成临床的误诊。     

脓毒症患者血清淀粉酶升高的临床意义

目的 探讨脓毒症患者血清淀粉酶升高的临床意义及其与预后的关系.方法 80例ICU脓毒症(APACHE Ⅱ评分>30分)患者,根据血清淀粉酶水平分为血清淀粉酶升高组(Ⅰ组)和正常组(Ⅱ组),比较两组血白细胞总数、低血压发生率、心律失常发生率、机械通气使用率、住院时间及病死率的变化.结果 80例脓毒症患者中,18例(22.5%)血清淀粉酶升高,胰腺损伤(血清淀粉酶≥400 U/L)发生率为11.3%.血清淀粉酶升高组低血压发生率、心律失常发生率、机械通气使用率、ICU住院时间、病死率明显高于血清淀粉酶正常组.血清淀粉酶水平愈高,预后愈差.血清淀粉酶≥400 U/L,病死率明显增高(P<0.01).结论 对脓毒症患者应注意血清淀粉酶的变化,血清淀粉酶可能是反映病情变化、判定顸后的一个重要指标.     

危重病患者血清淀粉酶升高的意义

目的研究危重病患者血清淀粉酶升高的临床意义及其与预后的关系。方法回顾性总结了125例急诊ICU危重病(APACHEⅡ评分＞29分)患者,根据血清淀粉酶水平分为血清淀粉酶升高组和正常组,比较两组血白细胞总数、低血压发生率、心力衰竭发生率、心律失常发生率、机械通气使用率、住院时间及病死率的变化。结果125例危重病患者中,28例(22．4%)血清淀粉酶升高,胰腺损伤(血清淀粉酶≥400 U/L)发生率为13．6%。血清淀粉酶升高组低血压发生率、心力衰竭发生率、心律失常发生率、机械通气使用率、ICU住院时间、病死率明显高于血清淀粉酶正常组。血清淀粉酶水平愈高,预后愈差。血清淀粉酶≥400 U/L,病死率明显增高(P＜0．01)。结论对危重病患者应注意血清淀粉酶的变化,血清淀粉酶可能是反映病情变化、判定预后的一个重要指标。     

血淀粉酶与血酸度的相关性研究

目的血清淀粉酶是一项常用的生化指标,对急性胰腺炎等疾病的诊断有重要价值,其测定值的准确性受实验底物、实验质控方法等多种因素的影响。本研究探讨作为酶动力学重要因素的血酸碱度对血清淀粉酶的影响。方法100例受检者,50例为胰腺炎,50例其他疾病病人。同时检测血淀粉酶及血pH值。淀粉酶检测应用日立770生化仪、方法为速率法;血pH检测为美国产ISTA血气分析仪、方法为干式法。资料用SPSS10.0软件进行统计分析。结果胰腺炎组酸中毒者的淀粉酶值与pH正常的患者比较差异无统计学意义（P=0.057）。胰腺炎组血淀粉酶与血pH值相关性检验相关性（r=-0.261,P=0.067）。其他疾病组pH值小于7.35与pH值正常者的淀粉酶值比较差异无统计学意义（P=0.072）。非胰腺炎组血淀粉酶与血pH值相关性检验无相关性（R=-0.477,P=0.47）。结论血清淀粉酶与血pH值相关性。