针对嗜麦芽糖拜瑞特氏菌白细胞毒素基因的实时PCR检测方法的建立与验证
Development and validation of a real-time PCR specific for the leukotoxin gene of Bibersteinia trehalosi.
作者信息
Killion Halcyon J, Edwards William, Jennings-Gaines Jessica, Wood Mary, Fox Karen, Sondgeroth Kerry
机构信息
Wildlife Health Laboratory (Killion, Edwards, Jennings-Gaines, Wood), Veterinary Services, Wyoming Game and Fish Department, Laramie, WY.
Department of Veterinary Sciences, University of Wyoming, Laramie, WY (Sondgeroth).
出版信息
J Vet Diagn Invest. 2018 Jul;30(4):589-592. doi: 10.1177/1040638717753497. Epub 2018 Mar 30.
A real-time PCR assay for the leukotoxin gene of Bibersteinia trehalosi was developed and validated to better identify this pathogen, which is a cause of respiratory disease in bighorn sheep. The specificity of the PCR primers was evaluated with DNA from 59 known isolates of the Pasteurellaceae family. For validation, 162 field samples were compared using both the new assay and an indirect method using 2 sets of published protocols. The real-time PCR assay was found to be specific for the leukotoxin gene of B. trehalosi and provides a rapid and direct approach for detecting leukotoxin-producing forms of this organism from samples containing mixed species of leukotoxin-positive Pasteurellaceae.
开发并验证了一种用于检测海藻糖巴氏杆菌白细胞毒素基因的实时PCR检测方法，以更好地鉴定这种病原体，它是大角羊呼吸道疾病的病因。用来自巴斯德菌科59种已知分离株的DNA评估了PCR引物的特异性。为了进行验证，使用新检测方法和使用2套已发表方案的间接方法对162份野外样本进行了比较。发现实时PCR检测方法对海藻糖巴氏杆菌的白细胞毒素基因具有特异性，并为从含有白细胞毒素阳性巴斯德菌科混合物种的样本中检测该生物体产生白细胞毒素的形式提供了一种快速直接的方法。
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