S. Razo, P. Galushka, Y. A. Varitsev, A. Zherdev, I. Safenkova, E. Pakina, B. Dzantiev
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引用次数: 0
摘要
马铃薯黑腿病是马铃薯最重要的细菌性病害之一。这种疾病在俄罗斯境内迅速蔓延,需要新的有效诊断工具,以便及时发现感染。为了解决这一问题,获得了Dickeya属的特异性抗血清。从抗血清中分离出的多克隆抗体对Dickeya spp.主要种(D. solani, D. dianthicola, D. chrysanthemum, D. dadantii, D. paradisiaca)具有高亲和力。酶联免疫吸附测定(ELISA)和侧流免疫测定(LFIA)测试系统已经基于获得的特异性和高亲和力抗体开发出来。ELISA法检测限为0.8 105个细胞/mL、2 104个细胞/mL。LFIA适用于非实验室条件,solani的检出限为2 105个细胞/mL,分析时间为15分钟。在检测马铃薯种子材料时,LFIA检测系统的ELISA检测结果75%为阳性,100%为阴性。
Development of new immunoanalytical test systems for diagnostics of potato blackleg caused by Dickeya spp. bacteria
Potato blackleg caused by Dickeya spp. bacteria is one of the most important bacterial diseases of potatoes. The rapid spread of this disease in the territory of Russia requires new effective diagnostic tools for the timely detection of infection. To solve this problem, antisera specific to Dickeya spp. were obtained. Polyclonal antibodies isolated from antisera have shown high affinity for the main species of Dickeya spp. ( D. solani, D. dianthicola, D. chrysanthemi, D. dadantii, D. paradisiaca ). Enzyme linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) test systems have been developed based on specific and high affinity antibodies that were obtained. For ELISA, the detection limit was 0.8 105 cells/mL for D. solani and 2 104 cells/mL for D. dianthicola . For LFIA, suitable for use in non-laboratory conditions, the detection limit of D. solani was 2 105 cells/mL and the analysis time was 15 minutes. When testing potato seed material, LFIA test system confirmed positive results of ELISA determination in 75 % of samples, and negative - in 100 % of samples.