Gel penetrate-blotted immunobinding assay, a novel method for serological detection of Fusarium spp. in soil
Gel penetrate-blotted immunobinding assay, a novel method for serological detection of Fusarium spp. in soil
复制标题
凝胶渗透印迹免疫结合测定，一种镰刀菌血清学检测的新方法。
DOI：
10.1584/jpestics.22.321
复制
发表时间：
1997
影响因子：
2.4
通讯作者：
I. Yamaguchi
中科院分区：
文献类型：
--
作者：
T. Arie;Y. Hayashi;K. Yoneyama;I. Yamaguchi
一种新的镰刀菌血清学检测方法。在土壤中建立。在一片硝酸纤维素膜上，覆盖约Imm厚的琼脂凝胶层，并将土壤样品悬浮液铺展在凝胶上。培养10至20小时后，样品中的真菌穿过凝胶层生长并到达膜，将土壤颗粒留在凝胶表面上。回收用真菌印迹的膜，并用抗镰刀菌属的单克隆抗体AP 19 -2处理，用碱性磷酸酶标记的第二抗体，最后是底物溶液。用该方法对镰刀菌侵染的土壤样品进行了免疫结合试验，结果表明该方法能在膜上显示阳性反应。我们将该方法命名为凝胶渗透印迹免疫结合试验（GP-IBA）。该方法可在24小时内完成，检测限约为103个分生孢子/g土壤。该方法被认为适用于检测镰刀菌属。在土壤中。
A new serological method for the detection of Fusarium spp. in soil was established. On a sheet of nitrocellulose membrane, about I mm thick layer of agar gel was overlaid, and soil sample suspension was spread over the gel. After 10 to 20 hr of incubation, fungi in the sample grew through the gel-layer and reached the membrane leaving the soil particles on the gel surface. The membrane blotted with the fungi was recovered, and was treated with a monoclonal antibody AP19-2 against Fusarium spp., secondary antibody labeled with alkaline phosphatase, and finally the substrate solution. The samples of Fusarium-infested soil successfully showed positive reaction on the membrane by this immunobinding assay (IBA). We named the method as gel penetrate-blotted immunobinding essay (GP-IBA). As the series of assay procedures can be completed within 24 hr and the detection limit is estimated to be about 10 3 conidia/g soil. this method is considered to be suitable for detection of Fusarium spp. in soil.