Instituto Valenciano de Investigaciones Agrarias, Moncada, Valencia, Spain.
Appl Environ Microbiol. 1994 Jun;60(6):2076-85. doi: 10.1128/aem.60.6.2076-2085.1994.
Seven monoclonal antibodies (MAbs) to Erwinia carotovora subsp. atroseptica have been produced. One, called 4G4, reacted with high specificity for serogroup I of E. carotovora subsp. atroseptica, the most common serogroup on potato tubers in different serological assays. Eighty-six strains belonging to different E. carotovora subsp. atroseptica serogroups were assayed. Some strains of serogroup XXII also reacted positively. No cross-reactions were observed against other species of plant pathogenic bacteria or 162 saprophytic bacteria from potato tubers. Only one strain of E. chrysanthemi from potato cross-reacted. A comparison of several serological techniques to detect E. carotovora subsp. atroseptica on potato tubers was performed with MAb 4G4 or polyclonal antibodies. The organism was extracted directly from potato peels of artificially inoculated tubers by soaking or selective enrichment under anaerobiosis in a medium with polypectate. MAb 4G4 was able to detect specifically 240 E. carotovora subsp. atroseptica cells per ml by indirect immunofluorescence and immunofluorescence colony staining and after soaking by ELISA-DAS (double-antibody sandwich enzyme-linked immunosorbent assay) after enrichment. The same amount of cells was detected by using immunolectrotransfer with polyclonal antibodies, and E. carotovora subsp. atroseptica and subsp. carotovora were distinguished by the latter technique. ELISA-DAS using MAb 4G4 with an enrichment step also efficiently detected E. carotovora subsp. atroseptica in naturally infected tubers and plants.
已经制备了 7 株针对胡萝卜软腐果胶杆菌亚种胡萝卜软腐亚种的单克隆抗体 (MAb)。一种称为 4G4 的抗体与胡萝卜软腐果胶杆菌亚种 I 血清型具有高度特异性反应，这是不同血清学检测中马铃薯块茎上最常见的血清型。对 86 株属于不同胡萝卜软腐果胶杆菌亚种胡萝卜软腐亚种血清型的菌株进行了检测。一些 XXII 血清型的菌株也呈阳性反应。对其他植物病原细菌或来自马铃薯块茎的 162 株腐生细菌没有交叉反应。只有一株来自马铃薯的菊欧文氏菌发生了交叉反应。使用 MAb 4G4 或多克隆抗体对几种检测马铃薯块茎上胡萝卜软腐果胶杆菌亚种胡萝卜软腐亚种的血清学技术进行了比较。将病原体直接从人工接种的马铃薯皮中提取出来，方法是在富含聚半乳糖酸盐的培养基中进行厌氧浸泡或选择性富集。通过间接免疫荧光和免疫荧光菌落染色，以及富集后的 ELISA-DAS（双抗体夹心酶联免疫吸附试验），MAb 4G4 能够特异性地检测到每毫升 240 个胡萝卜软腐果胶杆菌亚种胡萝卜软腐亚种细胞。使用多克隆抗体进行免疫电转移也可以检测到相同数量的细胞，并且后一种技术可以区分胡萝卜软腐果胶杆菌亚种和胡萝卜软腐亚种。使用 MAb 4G4 进行 ELISA-DAS 并进行富集步骤也可以有效地检测到自然感染的块茎和植物中的胡萝卜软腐果胶杆菌亚种胡萝卜软腐亚种。
