A novel assay system for myeloperoxidase activity in whole saliva
A novel assay system for myeloperoxidase activity in whole saliva
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DOI：
10.1016/j.clinbiochem.2007.12.025
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发表时间：
2008-05-01
影响因子：
2.8
通讯作者：
Izumi, Hiroshi
中科院分区：
文献类型：
--
作者：
Sakamoto, Wataru;Fujii, Yoshihiro;Izumi, Hiroshi
目的：应用新型测定系统直接测量全唾液中的 MPO（髓过氧化物酶）活性。设计和方法：该测定系统在氨苯砜（4,4'-二氨基二苯砜）存在下采用 3,3'-二氨基联苯胺 (DAB) 和愈创木酚的新型敏感底物，使用原始的“三明治”测试盘（DEAE-纤维素纸和纤维素色谱法）测定全唾液中的 MPO 活性纸）。将唾液（0.1 mL）直接涂在夹心测试盘上，然后添加 0.1 mL 含有 1 mM 氨苯砜的 0.3 M Tris-HCl 缓冲液（pH 7.5）的底物溶液。室温孵育30分钟后，用光学分析仪在460 nm处测量测试盘上的吸光度。结果：该检测系统能够区分全混合唾液中的MPO和唾液过氧化物酶，并且灵敏、简便且廉价。检测结果显示，患有牙周病的受试者的全部唾液中的 MPO 活性显着高于健康受试者的唾液中的 MPO 活性。 MPO 活性与龈下袋探诊深度也呈显着正相关（r=0.736，P
Objectives: The application of a novel assay system for the direct measurement of MPO (myeloperoxidase) activity in whole saliva.Design and methods: The assay system employs a novel sensitive substrate from 3,3'-diaminobenzidine (DAB) and guaiacol in the presence of dapsone (4,4'-diaminodiphenylsulfone) to determine MPO activity in whole saliva using an original "sandwich" test-disk (DEAE-cellulose paper and cellulose chromatography paper). The saliva (0.1 mL) was directly applied to the sandwich test-disk, and then 0.1 mL of the substrate solution containing 1 mM dapsone in 0.3 M Tris-HCI buffer (pH 7.5) was added. After incubation for 30 min at room temperature, absorbance on the test-disk was measured at 460 nm with an optical analyzer.Results: The assay system was shown to distinguish MPO from salivary peroxidase in whole mixed saliva and was sensitive, easy and cheap. The assays revealed that MPO activity in whole saliva from subjects with periodontal disease was significantly higher than in saliva from healthy subjects. There was also a significant positive correlation between MPO activity and the probing depth of subgingival pockets (r=0.736, P