科研动态

为建立一种检测兔源肺炎克雷伯氏菌（Klebsiella pneumoniae）重组酶聚合酶扩增方法（recombinasepolymerase amplification，RPA），根据肺炎克雷伯氏菌的phoE基因保守序列设计引物，扩增片段大小为277 bp，并对反应条件进一步优化，最终建立了适宜于快速准确检测兔源肺炎克雷伯氏菌的重组酶聚合酶等温扩增方法。该方法可特异性检测出兔源肺炎克雷伯氏菌，最低检出细菌量为8.3×101 CFU/mL，灵敏度比传统PCR高100倍。本研究建立的肺炎克雷伯氏菌RPA检测方法特异性强、操作简单，从而为生产中的兔源肺炎克雷伯氏菌现场快速检测提供了一种新方法。

Establishmentof a RPA Assay for Detecting Klebsiella pneumoniae of Rabbit Origin

In order to establish a recombinasepolymerase amplification（RPA）assay for detecting Klebsiella pneumonia of rabbit origin，a pair of primers were designed based on the conservative sequenceof PhoE gene of Klebsiella pneumoniae，and the fragmentwith the length of 277 bp was amplified，then thereaction conditions were further optimized，finally，the RPA assay was established，by which，the Klebsiella pneumoniae of rabbit origin could be specificallydetected with the minimum bacteria of 8.3×101 CFU/mL，and its sensitivity was 100 times higher than that of traditionalPCR. Therefore，the method established in the study hadcharacteristics of strong specificity and simple operation，which would provide a new method for rapid detection of Klebsiellapneumoniae of rabbit origin.

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