Cooray R, Petersson C G, Moberg L, Båge R
National Veterinary Institute, Uppsala, Sweden.
Zentralbl Veterinarmed B. 1995 Oct;42(8):481-91. doi: 10.1111/j.1439-0450.1995.tb00739.x.
The heme enzyme myeloperoxidase (MPO), with a spectral A430/A280 ratio > 0.78, was purified from isolated bovine neutrophils. Using highly specific anti-MPO monoclonal and anti-MPO polyclonal antibodies raised against MPO, a specific and sensitive double-antibody enzyme-linked immunosorbant assay (ELISA) was developed to measure bovine MPO in serum and neutrophil extracts. The ELISA shows good precision and accuracy, with intra- and interassay coefficients of variation of < 10% for MPO concentrations ranging from 0.5 to 50 ng/ml. The accuracy of the ELISA for measuring MPO in bovine serum was further confirmed by the similarity between the standard curve and curves obtained with successive dilutions of MPO-rich serum samples. The mean analytical recovery of MPO from serum was approximately 90%. Long delays between blood sampling and serum preparation were found to affect the level of MPO in the serum. Mean MPO values in the serum of healthy adult cows were 6.5 ng/ml, with a range of 3.5-15.3 ng/ml. In dairy cows with acute mastitis, mean serum MPO values were approximately 30 ng/ml, with a range of 6.0-59.6 ng/ml, and the elevation was markedly higher than the normal values (P = 0.0001). In isolated neutrophils from healthy cattle, MPO concentrations were found to be 7 x 10(-4) ng, with a range of 6.5-8.3 x 10(-4) ng/neutrophil. The ELISA was used to study the distribution of MPO in the bovine neutrophil granules; it was found to be localized to one distinct compartment.
血红素酶髓过氧化物酶（MPO）的光谱A430/A280比值>0.78，是从分离的牛中性粒细胞中纯化得到的。利用针对MPO产生的高度特异性抗MPO单克隆抗体和抗MPO多克隆抗体，开发了一种特异性和敏感性高的双抗体酶联免疫吸附测定法（ELISA），用于测定血清和中性粒细胞提取物中的牛MPO。该ELISA显示出良好的精密度和准确性，对于浓度范围为0.5至50 ng/ml的MPO，批内和批间变异系数<10%。通过标准曲线与富含MPO的血清样品连续稀释所得曲线之间的相似性，进一步证实了ELISA测定牛血清中MPO的准确性。MPO从血清中的平均分析回收率约为90%。发现采血和血清制备之间的长时间延迟会影响血清中MPO的水平。健康成年奶牛血清中的平均MPO值为6.5 ng/ml，范围为3.5 - 15.3 ng/ml。在患有急性乳腺炎的奶牛中，血清MPO的平均值约为30 ng/ml，范围为6.0 - 59.6 ng/ml，且升高明显高于正常值（P = 0.0001）。在来自健康牛的分离中性粒细胞中，发现MPO浓度为7×10(-4) ng，范围为6.5 - 8.3×10(-4) ng/中性粒细胞。该ELISA用于研究MPO在牛中性粒细胞颗粒中的分布；发现它定位于一个独特的区室。
