微量琼脂糖弥散法抗菌试验的建立及应用

Agrose Gel Diffusion Assays for Endogenous An ti bacterial Polypeptides

作者建立了两种微量而敏感的抗菌试验，可用于鉴定存在于细胞及组织中的抗菌蛋白或多肽。①琼脂糖弥散法，用其检测抗菌蛋白的抗菌活性，具有操作简便、灵敏度高、样品用量少（ｎｇ水平）、可同时检测多个样品及定量检测等优点；②电泳凝胶琼脂糖弥散法，该法在微量样品来经纯化的情况下，可直接确定存在于ＰＡＧＥ凝胶中抗菌蛋白条带。应用这两种方法作者首次鉴定出大鼠膀胱粘膜、人膀胱粘膜以及大鼠子宫内膜存在抗菌蛋白。

Two sensitive metheds for identifying the endogenous antibecterlal polypeptides were established in ourlaberatory. One methed uses an agrose gel radlal diffusion assay for bactericidal activity,This methed allows muItiplesamples to be tested。The other methed applys a gel overlay technique to identify antibecterial polypeptides in samples sub-jected to acid-urea polyacrylamide gel electrophoresis.When E。coli ML-35p was used as a test organism in the agreee gelradial diffusion assay,abeut 30 ng of human or rabbit defensins (HNP or NP-l) in 5ul of sample was routinely detect-ed. By the two methods。 we identified antibecteriaI polypoptides in the mucosa of rat. rabbit.and human bladders. andin the rat uterus wall.