甘薯青枯病潜症苗的快速检测

Rapid Detection of Pseudomonas Solanacearum in Sympton-latent Seedings of Sweet Potato

建立6株分泌抗甘薯青枯病菌特异性单克隆抗体的杂交瘤细胞。该单抗对甘薯青枯病菌以及可侵染甘薯的7个参试青枯病菌呈阳性反应;与参试的其它假单胞杆菌、其它属的检测菌株及著苗上的各种杂菌无交叉反应。ELISA法测定培养液抗体滴度为10~2—10~3;腹水抗体滴度为10~5—10~6。检出抗原灵敏度为每ml11.5μl蛋白。杂交瘤细胞及其培养液抗体保存3年以上均保持原有特性。6株单抗对我省分离的3个致病型的甘薯青枯病菌全部产生阳性反应,覆盖面广。ELISA间接法检测人工接种的潜症薯苗,可准确地检出带菌,检出率达100％。对119份病田无症苗进行检测,其中51份阳性反应,68份阴性反应。通过单抗检测与接种检测的比较试验,说明单抗检测的敏感性和特异性都高于接种法,同时快速简便。

Six hybridoma cell lines secreting monoclonal antibodies (McAbs) against the pathogen of Pseudomonas solanacearum from sweet potato have been produced. The McAbs showed positive reactions to 7 sweet-potato-infected strains, and negative to other bacteria of Pseudomonas and genus else, and various bacteria infected seedings of sweet potato detected in the test. The ELISA liters of the culture supernatanls and the ascites antibodies were respectively 10^(2)-10^(3) and 10^(5)-10^(6).The sensitivity of antigen detected was 11. 5ng/ml (protein concentration) . The original characters of the hybridoma cell and culture supernatants antibodies could be kept over three years. Six antibodies showed positive to all of 3 pathogenicity types of Pseudomonas solanacearum from sweet potato in Fujian province. The inoculated samples could be detected out by the indirect ELISA at the rate of 100%. In 119 portions of symptonlatent seedlings collected from field cases, 51 and 68 were respectively positive and negative. To compare with the antibody and inoculation detection, the result showed the sensitivity and specificity of antibody detection were more height than that of inoculation detection, and more rapid and handy.