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首页> 外文期刊>Phytopathologia Mediterranea >A real-time PCR quantitative detection assay for Pseudomonas savastanoi pv. nerii in Nerium oleander
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A real-time PCR quantitative detection assay for Pseudomonas savastanoi pv. nerii in Nerium oleander

机译：实时荧光定量定量检测法对假单胞菌PV。夹竹桃夹竹桃

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A real-time PCR assay based on TaqMan chemistry was developed for the detection of Pseudomonas savastanoi pathovars that cause bacterial knot disease on different plant species. Primers and probe sequences were based on the iaaL gene coding for (indole-3-acetyl)-L-lysine synthetase and previously used in conventional PCR tests. Assay specificity was tested with an extended range of strains of P. savastanoi from eight hosts, with 13 other Pseudomonas spp., and with other microorganisms naturally occurring on or in oleander plants. A pure culture cell suspension was quantified over a seven log concentration range (10(8) to 10(2) cfu ml(-1)). Different protocols were developed for the detection and quantification of P. savastanoi pv. nerii from symptomatic and asymptomatic oleander plants. A 24-h bacterial enrichment step either on PVF-1 or OKA-M broth improved the sensitivity of the assay, making it suitable to screen planting material for latent infections.
机译：开发了一种基于TaqMan化学的实时PCR检测试剂盒，用于检测导致不同植物物种上的细菌性结节病的假单胞菌假单胞菌病。引物和探针序列基于编码（吲哚-3-乙酰基）-L-赖氨酸合成酶的iaaL基因，以前用于常规PCR测试。用来自八种宿主，其他十三种假单胞菌属和夹竹桃植物上或之中天然存在的其他微生物的大范围沙瓦氏疟原虫菌株测试了测定特异性。在七个对数浓度范围（10（8）至10（2）cfu ml（-1））上对纯培养细胞悬浮液进行定量。开发了用于检测和定量沙门氏菌PV的不同方案。有症状和无症状夹竹桃植物的nerii。在PVF-1或OKA-M肉汤上进行的24小时细菌富集步骤提高了测定的灵敏度，使其适合筛选潜在感染的种植材料。

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