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酶抗体法于1967年Nakalle氏首先提出的。它比萤光抗体法有更多的优点,还能进行光学显微镜与电子显微镜的观察。本文作者采用了酶抗体法进行了牙本质龋的研究。酶抗体液的制作,是用变形链球菌与金黄色葡萄球菌做为抗原,去免疫家兔,所得到的抗血清用过氧化物酶进行标记,染色全部使用酶抗体直接法进行。过氧化物酶的检出,是选用三种基质做为底物:即3.3’二氨基联苯胺(简称DAB,反应物呈黄褐色)、α—萘酚—二苯氧芑胺(反应物呈粉红色)、4—氯—1萘酚(反应物呈灰蓝色),可顺序进行三重染色,亦可采用DAB与其中另一种染色进行二重染色。
The enzyme antibody method was first proposed by Nakalle in 1967. It has more advantages than the fluorescent antibody method, but also for optical and electron microscopy. The authors used the enzyme-antibody method to study dentine caries. Enzyme antibody solution was made by using Streptococcus mutans and Staphylococcus aureus as antigens to immunize rabbits. The obtained antiserum was labeled with peroxidase, and the staining was performed by enzyme antibody direct method. Peroxidase was detected using three substrates as substrate: 3.3 'diaminobenzidine (DAB, the reaction was yellowish brown), α-naphthol-diphenoxylamine (reactant was Pink), 4-chloro-1-naphthol (reactants are gray-blue), triple-stained sequentially, and double-stained with DAB and one of the other.