The Food and Environment Research Agency, Sand Hutton, York YO41 1LZ, United Kingdom.
J Microbiol Methods. 2010 May;81(2):116-20. doi: 10.1016/j.mimet.2010.02.006. Epub 2010 Feb 18.
In a direct comparison with established methods for Phytophthora ramorum detection (isolation followed by morphological identification, or conventional DNA extraction followed by TaqMan real-time PCR) a rapid, simplified detection method in which membranes of lateral flow devices (LFDs) are added directly to TaqMan real-time PCR reactions was used to test 202 plant samples collected by plant health inspectors in the field. P. ramorum prevalence within the 202 samples was approximately 40% according to routine testing by isolation or TaqMan real-time PCR. The diagnostic sensitivity and specificity of the rapid detection method were 96.3% and 91.2%, respectively. This method can be used in conjunction with Phytophthora spp. lateral flow devices to reduce the number of samples requiring testing using more laborious conventional methods. The effect of combining prescreening for Phytophthora spp. with P. ramorum-specific tests is discussed in terms of the positive and negative predictive values of species-specific detection when testing samples collected in different inspection scenarios.
与已建立的腐霉属 ramorum 检测方法（分离后进行形态鉴定，或常规 DNA 提取后进行 TaqMan 实时 PCR）进行直接比较，使用一种快速、简化的检测方法，即将侧向流装置（LFD）的膜直接添加到 TaqMan 实时 PCR 反应中，对植物健康检查员在现场采集的 202 个植物样本进行了测试。根据常规的分离或 TaqMan 实时 PCR 检测，202 个样本中腐霉属 ramorum 的流行率约为 40%。快速检测方法的诊断灵敏度和特异性分别为 96.3%和 91.2%。该方法可与腐霉属侧向流装置结合使用，以减少需要使用更繁琐的常规方法进行测试的样本数量。结合腐霉属的预筛选与腐霉属 ramorum 特异性测试的效果，根据在不同检查场景中收集的样本进行物种特异性检测的阳性和阴性预测值进行了讨论。
