免疫凝聚试纸条和TaqMan探针实时荧光PCR检测西瓜细菌性果斑病菌比较研究

 以西瓜细菌性果斑病菌(Acidovorax avenae subsp.citrulli)菌悬液和田间采集的病组织为试材,研究了免疫凝聚试纸条和实时荧光PCR技术检测的灵敏度和适应性。结果表明,免疫凝聚试纸条检测灵敏度为10⁶ cfu/mL,具有简便、快速、易操作特点,适用于田间快速检测和病害诊断;TaqMan探针实时荧光PCR检测灵敏度达10^3~4 cfu/mL,比传统PCR检测灵敏度(10⁵ cfu/mL)提高了10~100倍,且不需要琼脂糖凝胶电泳、溴化乙锭染色和Southern杂交。但需要昂贵的仪器和试剂,适用于室内检测及相关研究。

Comparison of immunoStrip and real-time fluorescent PCR (TaqMan)for detection of Acidovorax avenae subsp. citrulli, the causal agent of bacterial fruit blotch of watermelon

To assess the threshold and efficiency of immunoStrip and real-time fluorescent PCR for detection of Acidovorax avenae subsp.citrulli in cell suspension from pure culture and in extracts from naturally infested or infected tissues,a comparative study was conducted.The results showed that immunoStrip testing,with threshold 10~ 6 cfu/mL,was simple,fast and easy to handle,which adapted for fast detection and diagnosis in field.Compared with classical-PCR threshold 10~ 5 cfu/mL,real-time fluorescent PCR TaqMan didn't require agarose gel electrophoresis,ethidium bromide staining,and Southern blotting.Moreover,it had a much higher sensitivity,accounting for 10~ 3-4 cfu/mL,but required expensive instrument,and was adapted to lab testing and basal research.