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当前位置: 首页 > 文章 > 应用DTBIA技术快速检测柑橘黄龙病菌研究 园艺学报 2018 (8) 1605-1612
Position: Home > Articles > Research on the Fast Detection of Candidatus Liberibacter Asiaticus with DTBIA Acta Horticulturae Sinica 2018 (8) 1605-1612

应用DTBIA技术快速检测柑橘黄龙病菌研究

作  者：
丁芳;王清廉;徐国亮;周佳乐;曾继吾;严翔;夏长秀;郭俊;刘永忠;洪霓;王国平;彭抒昂;John S Hartung
关键词：
柑橘;黄龙病菌;快速检测;DTBIA;多克隆抗体;特异性
摘  要：
柑橘黄龙病病原菌为韧皮部限制性寄生菌Candidatus Liberibacter,迄今为止尚未成功获得离体纯培养。因此基于特异性抗体的高通量血清学检测技术迄今尚未开发。本实验室前期研究利用异源表达重组黄龙病菌外膜蛋白Omp_(CLas)免疫动物,制备并获得了特异性多克隆抗体(Anti-Omp_(CLas)-Pab)。本研究利用直接组织印迹免疫法(Direct tissue blot immunoassay,DTBIA)技术对获得的多克隆抗体的检测效果进行了初步评价,证实该抗体可以特异性识别中国不同地理来源的柑橘黄龙病菌分离物;柑橘叶柄、叶片中脉、枝条、根部、果梗和种子中均存在黄龙病菌,其中根部和种子中的病菌含量相对较低,枝条和叶片中脉中的含量较高。该多克隆抗体的获得为后续快速检测试剂盒或试纸条的研发奠定了基础。
译  名：
Research on the Fast Detection of Candidatus Liberibacter Asiaticus with DTBIA
作  者：
DING Fang;WANG Qinglian;XU Guoliang;ZHOU Jiale;ZENG Jiwu;YAN Xiang;XIA Changxiu;GUO Jun;LIU Yongzhong;HONG Ni;WANG Guoping;PENG Shu'ang;John S HARTUNG;Hubei Key Laboratory of Plant Pathology,Huazhong Agricultural University;Key Laboratory of Horticultural Plant Biology(Huazhong Agricultural University),Ministry of Education;State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University;Institute of Fruit Trees,Guangdong Academy of Agricultural Sciences;Ganzhou Citrus Research Institute;Institute of Tropical and Subtropical Cash Crops,Yunnan Academy of Agricultural Sciences;USDA-ARS-MPPL;
单  位：
Huazhong Agricultural University
关键词：
Citrus;;HLB bacteria;;fast detection;;DTBIA;;polyclonal antibody;;specificity
摘  要：
Citrus Huanglongbing(HLB)is caused by phloem-limited bacteria named Candidatus Liberibacters. Up to date,despite of all kinds of efforts to get pure culture,HLB bacteria still resist in vitro growth. Fast detection especially the high-throughput methods based on the specificity of serological methods has not yet been developed. In our previous work,we constructed fusion OmpCLas protein and heterogonous expressed to get antigen to produce corresponding polyclonal antibody(Pab). In the present study,we carried out experiment on the assessment of the detection efficiency of Anti-OmpCLas-Pab antibody with direct tissue blot immunoassay( DTIBA). Citrus samples collected from different geographical regions were tested. Different organs of petiole,midrib,stem,root,peduncale and seeds were all analyzed. Detection efficiency of both DTBIA and PCR were compared. Our result indicated:AntiOmpCLas-Pab could specifically recognize HLB bacteria in different isolates originated from different geographical areas in China,including samples from Jiangxi,Fujian,Yunnan,Guangdong and Hainan provinces. Tests with different tissues of petiole,leaf mid rib,stem,root,peduncale and seeds revealed the presence of HLB bacteria. However,less HLB bacteria were found in roots and seeds;higher titer was found in leaf mid-ribs and stems. Our results indicated that Anti-OmpCLas-Pab is a promising antibody for the fast detection of HLB bacteria which laid a basic foundation for the product development on the fast detection kits and dipsticks.

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