Olsson T, Sandstedt K, Holmberg O, Thore A
Biotechnol Appl Biochem. 1986 Oct;8(5):361-9.
The release of ATP from somatic cells in milk with the detergent Triton X-100 was optimized for assay with firefly luciferase. A small volume of milk (40 microliters) is added to 0.8 ml of 0.2% Triton X-100 in 100 mM Tris, 4 mm EDTA, pH 7.8. After approximately 1 min, 0.2 ml of luciferase reagent is added and the emission of light is measured in a luminometer. Results are calibrated with an ATP standard. This single method gave high yields of ATP from somatic cells in milk without interference from bacterial ATP. Extracts could be stored or transported prior to assay without deterioration of results. A close correlation was found between somatic cell count and ATP in milk samples collected at a farm as well as in milk samples from a cow with experimental mastitis. Results are promising for future use for diagnosis of mastitis but further work and field testing has to be done before it can be used on a wider scale.
为了用萤火虫荧光素酶进行检测，对用去污剂Triton X - 100从牛奶中的体细胞释放ATP的方法进行了优化。将少量牛奶（40微升）加入到0.8毫升含0.2% Triton X - 100的100 mM Tris、4 mM EDTA（pH 7.8）溶液中。大约1分钟后，加入0.2毫升荧光素酶试剂，并在发光计中测量发光情况。结果用ATP标准品进行校准。这种单一方法能从牛奶中的体细胞获得高产率的ATP，且不受细菌ATP的干扰。提取物在检测前可以储存或运输，结果不会变差。在农场采集的牛奶样本以及患有实验性乳腺炎奶牛的牛奶样本中，发现体细胞计数与ATP之间存在密切相关性。该结果对于乳腺炎的未来诊断很有前景，但在更广泛应用之前还需要进一步的研究和现场测试。
