水稻细菌性谷枯病菌和水稻白叶枯病菌荧光RPA快速检测方法的建立

Establishment of fluorescence RPA detection methods for Burkholderia glumae and Xanthomonas oryzae pv. oryzae

本研究分别针对我国进境植物检疫性有害生物水稻细菌性谷枯病菌及水稻白叶枯病菌建立了光RPA检测方法,并对其灵敏度、特异性以及对实际种子样本的检测能力进行了测试评价。结果表明,本研究建立的两种方法均能够在20 min内特异性检测到目标菌株,对水稻细菌性谷枯病菌(Burkholderia glumae)菌液的检测下限达到了11.4 CFU/反应,DNA检测下限达到了4.83×10^(-5)ng/反应;对水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae)菌液的检测下限达到了4.42 CFU/反应,DNA检测下限达到了3.83×10^(-4)ng/反应。两种方法均能够成功应用于带菌种子样品的快速检测。

Two fluorescence RPA methods for detection of quarantine bacterium Burkholderia glumae and Xanthomonas oryzae pv.oryzae were developed respectively.The sensitivity,specificity and the ability to detect actual seed samples of these two methods were evaluated.The detection results suggested that the target strains could all be detected within 20 min using the RPA methods established in this study.The detection threshold of Burkholderia glumae is 11.4 CFU/reaction for bacteria culture and 4.83×10^(-5)ng/reaction for DNA.While the detection threshold of Xanthomonas oryzae pv.oryzae is 4.42 CFU/reaction for bacteria culture and 3.83×10^(-4)ng/reaction for DNA.Both methods can be successfully applied to the rapid detection of infected seed samples.