| 摘要: |
| 选取不同浓度的柑橘溃疡病菌SOB培养液，利用载玻片进行固定、封闭、抗体结合、免疫荧光标记及荧光强度检测，以探讨免疫荧光检测技术在柑橘溃疡病菌快速检测方面的应用。结果表明，30 ℃下用BSA封闭2 h，30 ℃下抗体结合1 h，荧光抗体浓度为4 μg/mL，室温放置40 min后，在荧光显微镜下可以清楚看到绿色的柑橘溃疡病菌体。该方法操作简便，检测过程仅需4 h，抗原抗体反应特异性极强，检测灵敏度可达103cfu/mL。 |
| 关键词: 柑橘溃疡病菌 免疫荧光检测 植物检疫 |
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| 基金项目:湖北省出入境检验检疫局重点科技项目（HUBE2005-04） |
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| Study on detecting Xanthomonas axonopodis pv.citri by immunofluorescence |
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| Abstract: |
| Citrus bacterial canker disease,caused by Xanthomonas axonopodis pv.citri，is a serious quarantine disease of most citrus species and cultivars in many citrus-producing areas worldwide.The study tried to use a piece of glass to establish a simple and convenient immunofluorescent technique for a quick identification of Xanthomonas axonopodis pv.citri.Immunofluorescence experiments demonstrate that green Xanthomonas axonopodis pv.citri bacteria bodies can be seen explicitly under fluorescent microscope after enclosed by BSA under 30 ℃ for 2 hours,combined with antibody for 1 hours,with the fluorescent antibody concentration of 4 μg/mL in room temperature for 40 minutes while Xanthomonas oryzae pv.oryzae can not.The testing with immunofluorescence antibodies by this means is so simple that only a fluorescent microscope is needed and the whole testing process only takes 4 hours with an extremely strong specificity in antigen antibody reaction and the assessing delicacy of less than 103 cfu/mL.Therefore,this method meets the practical demands of the rapid Citrus bacterial canker quarantine identification quite well in export and import sites. |
| Key words: Xanthomonas axonopodis pv.citri IgG-FITC immunofluorescence detection plant quarantine |
