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根据香石竹细菌性萎蔫病菌基因组16S-23S rRNA保守序列,设计并合成了一对特异性引物和一条具有稳定点突变特异性探针,建立了对香石竹细菌性萎蔫病菌的TaqMan实时荧光PCR检测方法。除香石竹细菌性萎蔫病菌外,还对其他7种病原细菌菌株进行了荧光PCR检测。结果表明,只有香石竹细菌性萎蔫病菌产生荧光,其他病原细菌均没有荧光产生。与常规PCR相比,实时荧光PCR检测特异性强,灵敏度高,能检测到浓度为0.4 pg/μL的DNA,且能直接用于苗木等样品的检测,适合病害的快速诊断和口岸检验检疫应用。
According to the 16S-23S rRNA conserved sequence of the bacterial wilt pathogen of carnation, a pair of specific primers and a probe with a stable point mutation were designed and synthesized, and a TaqMan real-time PCR detection of the bacterial wilt disease of carnation method. In addition to carnation bacterial wilt disease bacteria, but also other seven kinds of pathogenic bacteria strains were detected by fluorescent PCR. The results showed that only Carnation bacterial wilt bacteria produce fluorescence, no other pathogenic bacteria fluorescence. Compared with conventional PCR, real-time PCR has the advantages of high specificity and high sensitivity, can detect DNA with a concentration of 0.4 pg / μL, and can be directly used for the detection of seedlings and other samples for rapid diagnosis of disease and quarantine applications at ports .