Citrus canker ( Xanthomonas citri subsp. citri ) is an important quarantine disease. Previous studies showed that loop-mediated isothermal amplification ( LAMP) can be used for rapid detection of suspected citrus can-ker samples. Based on a new conserved genome DNA sequence, we designed LAMP primers and established a LAMP detection method for detection of citrus canker samples in this study. The specificity of established LAMP detection method was consistent with that of regular PCR. However, this LAMP detection method could detect as low as 100 pg DNA of X. citri subsp. citri, and only 4 cells per microliter of X. citri, sensitivity of LAMP was 1250 times higher than that of the regular PCR. The LAMP assay could distinguish citrus canker from citrus scab, the two diseases eas-ily confused based on symptoms. Therefore, we suggest that LAMP detection method for citrus canker established in this study is simple, rapid, sensitive and specific, it could be used in laboratory with relatively simple conditions.%环介导等温扩增(loop-mediated isothermal amplification,LAMP)快速检测已用于基层植物检验检疫中的疑似样品检测诊断.柑橘溃疡病菌(Xanthomonas citri subsp.citri)是重要的检疫性病害,根据柑橘溃疡病菌基因组中一段保守序列设计LAMP引物,建立了柑橘溃疡病菌的LAMP检测方法.该LAMP检测方法只对柑橘溃疡病菌DNA和柑橘溃疡病病斑总DNA进行扩增,而对非靶标菌DNA和非靶标病斑总DNA不发生扩增,特异性与PCR一致.该LAMP检测方法最低可检出100 pg的纯柑橘溃疡病菌DNA和最低每微升4个细菌,灵敏度均比PCR高了1250倍.LAMP检测法能很好地区分症状极易混淆的柑橘溃疡病和疮痂病.该试验建立的柑橘溃疡病菌LAMP检测方法具有简单、快速、灵敏和特异等优点,可在条件相对简陋的实验室应用.
展开▼
