An immunoturbidimetric assay for rapid quantitative measurement of feline alpha-1-acid glycoprotein in serum and peritoneal fluid
An immunoturbidimetric assay for rapid quantitative measurement of feline alpha-1-acid glycoprotein in serum and peritoneal fluid
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DOI：
10.1111/j.1939-165x.2005.tb00058.x
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发表时间：
2005-12-01
影响因子：
1.2
通讯作者：
Eckersall, PD
中科院分区：
文献类型：
--
作者：
Bence, LM;Addie, DD;Eckersall, PD
背景：α-1-酸性糖蛋白（AGP）是一种急性时相蛋白，在感染和炎症条件下浓度增加。血清和腹腔液中的AGP浓度可能有助于诊断猫传染性腹膜炎（FIP），这是一种猫的致命疾病。目前，AGP可以通过放射免疫扩散（RID）测定来测量，这是耗时且困难的。目的：本研究的目的是建立一种快速的免疫比浊法测定猫血清和腹腔液中的AGP，并与RID的结果进行比较。研究方法：通过高氯酸沉淀和离子交换色谱法从患有FIP的猫的腹腔液池中纯化AGP，通过一组实验室检测确定，包括血清AGP浓度、白蛋白：球蛋白比和总蛋白浓度、抗冠状病毒抗体滴度和渗出液分析。将在完全弗氏佐剂和吐温20混合物中的纯化的AGP注射到绵羊中，并每月收集血液。使用经ELISA和Western印迹技术确认的抗AGP抗血清和来自患有FIP的猫的腹膜液池制备标准品。对猫腹腔液（n = 55）和血清（n = 59）的临床样本进行了AGP测定，并对免疫比浊法和RID法的结果进行了比较。结果：腹膜液（R-2 = 0.9259）和血清（R-2 = 0.9448）样本的方法之间具有显著相关性（P <0.001）。免疫比浊法的变异系数为
Background: Alpha-1-acid glycoprotein (AGP) is an acute phase protein that increases in concentration in infectious and inflammatory conditions. The serum and peritoneal fluid concentrations of AGP may be useful in the diagnosis of feline infectious peritonitis (FIP), a lethal disease of cats. Currently AGP can be measured by radioimmunodiffusion (RID) assays, which are time-consuming and difficult. Objectives: The objectives of this study were to develop a rapid immunoturbidimetric assay for measurement of AGP in feline serum and peritoneal fluid and to compare the results with those obtained by RID. Methods: AGP was purified by perchloric acid precipitation and ion-exchange chromatography from a pool of peritoneal fluid obtained from cats with FIP, as determined by a panel of laboratory tests, including serum AGP concentration, albumin: globulin ratio, and total protein concentration, anti-coronavirus antibody titers, and effusion analysis. The purified AGP in a complete Freund's adjuvant and Tween 20 mixture was injected into a sheep and blood was collected at monthly intervals. Anti-AGP antiserum, as confirmed by ELISA and Western blot techniques, and a pool of peritoneal fluid from cats with FIP were used to prepare standards. Clinical samples of feline peritoneal fluid (n = 55) and serum (n = 59) were assayed for AGP and results from the immunoturbidimetric and RID methods were compared. Results: Significant correlation (P < .001) was obtained between methods for both peritoneal fluid (R-2 = .9259) and serum (R-2 = .9448) samples. Coefficients of variation for the immunoturbidimetric method were