Development of a real-time PCR-based method for detection of Xylophilus ampelinus
Development of a real-time PCR-based method for detection of Xylophilus ampelinus
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DOI：
10.1111/j.1365-3059.2006.01487.x
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发表时间：
2007-02-01
期刊：
影响因子：
2.7
通讯作者：
Ravnikar, M.
中科院分区：
文献类型：
--
作者：
Dreo, T.;Gruden, K.;Ravnikar, M.
建立了葡萄白叶枯病菌的荧光定量PCR检测方法。与DNeasy植物微型试剂盒联用，X.从组织提取物中检测到大约100个细胞，超过现有巢式PCR方法的灵敏度至少十倍。在田间样品中，观察到获得的实时PCR循环阈值（Ct）值与X。在人工培养基上分离葡萄酒。从Ct值低于25的样品中成功分离。较低浓度的X。在实时荧光PCR中也可以可靠地检测到Ct值高达36的ampelinus。新开发的方法提供了一个可靠和灵敏的X。ampelinus，适合作为筛选试验，补充分离的媒体或其他方法，也可用于快速和特异性鉴定分离的菌落和X的相对定量。ampelinus细菌
A real-time PCR MGB-probe-based detection method specific to Xylophilus ampelinus, the cause of grapevine bacterial blight, was developed. Used in combination with the DNeasy plant mini kit, the sensitivity of X. ampelinus detection was approximately 100 cells from tissue extracts, surpassing the sensitivity of an existing nested PCR method at least tenfold. In field samples a high correlation was observed between real-time PCR cycle threshold (Ct) values obtained and X. ampelinus isolation on artificial media. Isolation was successful from samples with Ct values below 25. Lower concentrations of X. ampelinus, with Ct values up to 36, could also be reliably detected in real-time PCR. The newly developed method offers a reliable and sensitive test for X. ampelinus, suitable as a screening test, complementary to isolation on media or other methods, and could also be used for fast and specific identification of isolated colonies and for relative quantification of X. ampelinus bacteria.