Sensitivity Comparison of Immunocapture PCR and Classical PCR for Detection of Burkholderia glurnae

• 作者: Chen Lei Liu Hui Luo Jinyan Wu Hanbing
• 来源期刊: Plant Diseases and Pests(植物病虫害研究：英文版)
• 学科: 农学
• 关键词: PCR方法 检测效率 灵敏度 免疫 捕获 氟离子选择性电极 抗血清鉴定 PCR检测
• 年，卷（期）: 2015,（3）
• 页码范围: 5-9
• 页数: 5页
• 分类号: S858.315.3

摘要

The present study combined the fluoride ion-selective electrode( ISE) method with classical PCR to increase the detection efficiency of Burkholderia glumae. The four antisera of rabbit anti whole cell of B. glumae had been successfully obtained by polyclonal antibody technique,which were ASBg14,ASBg32,ASBg26 and ASBg04,respectively. The titers of four antisera reached 1∶ 32 in ODD test and 1∶ 5 120 in agglutination reaction. All antisera were identified to be qualified antibodies through combined method. The specificity of ASBg14 and ASBg32 was very high,but that of ASBg26 and ASBg04 was not very satisfying. The results showed that all tested strains of B. glumae produced 500 bp specific fragments by direct PCR and immunocapture PCR detection,while others showed negative PCR result. Comparing the sensitivity of these two detection methods,direct PCR could detect about 1 × 10~5 cfu / m L of bacterial suspension while immunocapture PCR could detect as few as 1 × 10~3 cfu / m L,10~2 times higher than direct PCR in sensitivity. In the detecting experiments of artificially inoculated seeds,at least two seeds were required in direct PCR,while only one seed was enough for immunocapture PCR detection.