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水貂绿脓杆菌环介导等温扩增快速检测方法的建立
引用本文:姜莉莉,樊兆斌,高文玉.水貂绿脓杆菌环介导等温扩增快速检测方法的建立[J].中国农学通报,2012,28(29):79-82.
作者姓名:姜莉莉  樊兆斌  高文玉
作者单位:辽宁医学院畜牧兽医学院,辽宁锦州,121001
基金项目:辽宁省教育厅项目(L2010259)
摘    要:为了利用环介导等温扩增(LAMP)技术建立水貂绿脓杆菌核酸快速检测方法。根据绿脓杆菌外毒素A基因240 bp片段保守序列设计一套LAMP特异性引物,对水貂绿脓杆菌基因组DNA进行LAMP扩增,通过浑浊度、SYBR Green荧光染料显色及电泳观察结果,并进行特异性和敏感性检验。结果表明:完成反应仅需要60 min(65℃),特异性强,最低检测限为1 ng/μL。该方法简单快捷、特异性强、灵敏度高,适合于基层使用,具有广泛的临床应用前景。

关 键 词:抗体  抗体  
收稿时间:2012/7/15 0:00:00
修稿时间:8/4/2012 12:00:00 AM

Rapid Detection for Mink Pseudomonas aeruginosa by Loop-mediated Isothermal Amplification
Jiang Lili , Fan Zhaobin , Gao Wenyu.Rapid Detection for Mink Pseudomonas aeruginosa by Loop-mediated Isothermal Amplification[J].Chinese Agricultural Science Bulletin,2012,28(29):79-82.
Authors:Jiang Lili  Fan Zhaobin  Gao Wenyu
Institution:(College of Animal Science and Veterinary Medicine,Liaoning Medical University,Jinzhou Liaoning 121001)
Abstract:In order to establish a rapid detection technique for mink Pseudomonas aeruginosa by Loop-mediated isothermal amplification.A set of specific primers were designed based on conserved region of PEA gene(Pseudomonas aeruginosa 240 bp),and amplified DNA for mink Pseudomonas aeruginosa by LAMP.The results were observed by turbidity,SYBR Green staining and agarose gel electrophoresis,meanwhile specificity and sensibility experiments were done too.The results showed that reaction was finished in 60 min,showed a high specificity and the detection limit was 1 ng/μL.The techniques were simple,rapid,high specificity and sensibility,so it could applied to base course and had comprehensive clinical prospect.
Keywords:mink  Pseudomonas aeruginosa  LAMP  establishment
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