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发明公开
CN105002280A 弧菌的荧光定量PCR检测方法 无效 - 驳回

弧菌的荧光定量PCR检测方法
摘要：
本发明提供了一种弧菌的荧光定量PCR检测方法，步骤包括：提取样本细菌培养后所得菌落的DNA，采用特异性引物对菌落的DNA进行荧光PCR扩增，所述特异性引物为以下引物对之一：toxR基因引物对：序列表SEQ ID No.1-2所示，flaA基因引物对：序列表SEQ ID No.3-4所示，pyrH基因引物对：序列表SEQ ID No.5-6所示。该方法特异性强、灵敏度高，并可进一步确定致病菌种类为河流弧菌、鳗弧菌或溶藻弧菌，可以检测出的最低浓度分别达到1.0×10-6nmol/L、1.0×10-7nmol/L、1.0×10-8nmol/L，检测方便快速准确，对防止海水养殖动物主要细菌性病害之一的弧菌病具有重要意义。
摘要（英）：

The invention provides a fluorogenic quantitative PCR detecting method of vibrios. The method comprises the steps that DNA of bacterial colonies obtained from culturing sample bacteria is extracted, fluorogenic PCR amplification is carried out on the DNA of the bacterial colonies through specific primers, and the specific primers are one of toxR gene primer pairs indicated by a sequence table SEQ ID No.1-2, flaA gene primer pairs indicated by a sequence table SEQ ID No.3-4, and pyrH gene primer pairs indicated by a sequence table SEQ ID No.5-6. The method is high in specificity and sensitivity, the pathogenic bacterium variety can be further determined as vibrio fluvialis or vibrio anguillarum or vibrio alginolyticus, the minimum concentration capable of being detected is 1.0*10<-6> nmol/L, 1.0*10<-7> nmol/L and 1.0*10<-8> nmol/L, detecting is convenient, fast and accurate, and great significance in preventing vibriosis, one of main bacterial diseases of maricultural animals, is achieved.

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