摘要
用浓度为10~9CFU/ml的X.c.pv.citri菌悬液免疫新西南雄兔,获取抗血清。采用硫酸铵盐析、透析脱盐、DEAE—纤维素层析法提纯兔抗X.c.pv.Citri IgG,用过碘酸钠法获得辣根过氧化物酶体与免抗X.c.pv.Citri IgG结合物,用双抗体夹心法检测柑桔溃疡病菌的灵敏度和特异性均比间接法高,柑桔叶片浸渍液经加热处理比不加热的检测灵敏度提高10倍左右。用双抗体夹心法检测了11个县(区)的129个不显症和可疑标样,有19个为阳性反应,结果与采集地实际情况相符。检测过程可在40小时内完成。表明用酶联免疫双抗体夹心法检测柑桔溃疡病菌灵敏度高,特异性强,准确、稳定、快速,且方便可行。
ELISA was evaluated for the detection of Xanthomonas campe-stris pv.citri (Hasse dye. The antisera were obtained from The New Zealand rabbits which were injected with X. c, pv. citri cell suspension (109CFU/mL) .The purified immuno globulin G (IgG) and horseradish Peroxidase conjugatse were prepared in laboratory. The sensitivities of indirectmethod-ELISA (IM-ELISA)and double sandwich method-ELISA(DSM-ELISA)were 103-4CFu/mL and 102-8CFU/ mL respectively. The sensitivity and specificity by DSM-ELISA were higher than by IM-ELISA
X.c.pv.citri was detected by the two methods in both heated(100℃ for 30min) and unheated cell suspensions of healthy leaf tissue extracts.The X. c. pv. citri specific reactiosn of heated preparations were generally greater than those of unheated ones.129 Symptomless citrus tissue samples from 11 ounties of Sichuan province, where the Citrus bacteral canker disease had been occurred, were detected by DSM-ELISA. 19 of them were positive reactions, X.c pv.citri cao be detected by ELISA technique.
出处
《西南农业学报》
CSCD
1993年第A00期55-60,共6页
Southwest China Journal of Agricultural Sciences
关键词
柑桔溃疡病
ELISA
柑桔
Xanthomonbs c.pv.citri Detection ELISA DSM-ELISA IM-ELISA
