Comparison of Protein Phosphatase Inhibition Assay with LC-MS/MS for Diagnosis of Microcystin Toxicosis in Veterinary Cases
Comparison of Protein Phosphatase Inhibition Assay with LC-MS/MS for Diagnosis of Microcystin Toxicosis in Veterinary Cases
复制标题
蛋白磷酸酶抑制测定与 LC-MS/MS 诊断兽医病例中微囊藻毒素中毒的比较
作者：
C. E. Moore;J. Juan;Yan;C. Gaskill;B. Puschner
微囊藻毒素是饮用水和休闲沃茨水中日益受到全球关注的急性肝毒素，对人类和动物的健康构成重大风险。由蓝藻产生的微囊藻毒素抑制丝氨酸/苏氨酸蛋白磷酸酶1（PP 1）。开发了一种使用对硝基苯基磷酸盐的具有成本效益的PP 1测定法，以快速评估水和瘤胃内容物样品。通过线性模型确定显著抑制，该模型将样品体积增加与暴露率与PP 1活性对照率的对数转换比进行比较。为了测试该模型在诊断病例调查中的有用性，对来自两个兽医病例的样本进行了测试。2013年8月，肯塔基州的两个池塘周围有15头牛死亡。一个池塘和三个测试瘤胃内容物有显着的PP 1抑制和微囊藻毒素-LR的检测水平，另一个池塘没有。2013年8月，一只狗在加州的清澈的湖水游泳后患上了致命的疾病。湖水样本收集后一个和四周的狗出现临床症状抑制PP 1活性。随后使用液相色谱-质谱（LC-MS/MS）分析检测到微囊藻毒素同系物-LR，-LA，-RR和-LF，但没有-YR。这些诊断研究说明了使用功能测定结合LC-MS/MS的优势。
Microcystins are acute hepatotoxins of increasing global concern in drinking and recreational waters and are a major health risk to humans and animals. Produced by cyanobacteria, microcystins inhibit serine/threonine protein phosphatase 1 (PP1). A cost-effective PP1 assay using p-nitrophenyl phosphate was developed to quickly assess water and rumen content samples. Significant inhibition was determined via a linear model, which compared increasing volumes of sample to the log-transformed ratio of the exposed rate over the control rate of PP1 activity. To test the usefulness of this model in diagnostic case investigations, samples from two veterinary cases were tested. In August 2013 fifteen cattle died around two ponds in Kentucky. While one pond and three tested rumen contents had significant PP1 inhibition and detectable levels of microcystin-LR, the other pond did not. In August 2013, a dog became fatally ill after swimming in Clear Lake, California. Lake water samples collected one and four weeks after the dog presented with clinical signs inhibited PP1 activity. Subsequent analysis using liquid chromatography-mass spectrometry (LC-MS/MS) detected microcystin congeners -LR, -LA, -RR and -LF but not -YR. These diagnostic investigations illustrate the advantages of using functional assays in combination with LC-MS/MS.
DOI：
10.1006/bbrc.1995.2605
发表时间：
1995-11-02
影响因子：
3.1
作者：
RUNNEGAR, M;BERNDT, N;ZHANG, LF
通讯作者：
ZHANG, LF
DOI：
10.1016/s0021-9258(17)45384-1
发表时间：
1990-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
R. Honkanen;J. Zwiller;R. Moore;S. L. Daily;B. Khatra;M. Dukelow;A. Boynton
通讯作者：
R. Honkanen;J. Zwiller;R. Moore;S. L. Daily;B. Khatra;M. Dukelow;A. Boynton
影响因子：
2.8
作者：
AN, JS;CARMICHAEL, WW
通讯作者：
CARMICHAEL, WW