中国兽医科学2018，Vol.48Issue(9)：1080-1085,6.DOI:10.16656/j.issn.1673-4696.2018.0150
副猪嗜血杆菌与猪链球菌2型双重LAMP检测方法的建立
Development of duplex LAMP assay for the simultaneously detection of Haemophilus parasuis and Streptococcus suis type 2
摘要
Abstract
To develop a duplex LAMP assay for the simultaneously detection of Haemophilus parasuis and Streptococcus suis 2, two sets of loop-mediated isothermal amplification (LAMP) primers were designed according to the inf B gene of H. parasuis and Cps2j gene of S. suis. The amplification products were then analyzed by electrophoresis,fluorescence observation and dissociation curves. The duplex LAMP assay could specifically detect H. parasuis and S. suis with a sensitivity that was about 100 times higher than the conventional PCR and the LAMP assay had no cross-reactions with the related pathogens. The exact pathogens could be also determined by the dissociation curve. A total of 29 clinical samples with respiratory disease were tested by the conventional PCR and the duplex LAMP and the positive rates were 17. 0％ and 31. 0％ ,respectively. These results demonstrated that the duplex LAMP assay was a useful technique for rapid and sensitive etiological diagnosis for H. parasuis and S. suis simultaneously and provided an effective method for the preliminary screening of clinical diseases.关键词
副猪嗜血杆菌/猪链球菌2型/双重LAMP/常规PCRKey words
Haemophilus parasuis/Streptococcus suis 2/duplex loop-mediated isothermal amplification/conventional PCR分类
农业科技引用本文复制引用
刘博婷,李娜,彭凌,郑娅莉,柯璐菲..副猪嗜血杆菌与猪链球菌2型双重LAMP检测方法的建立[J].中国兽医科学,2018,48(9):1080-1085,6.基金项目
广东省公益研究与能力建设专项资金资助项目(2014A020208140) （2014A020208140）
粤北生猪生产及疫病防控协同创新发展中心资助项目[粤教科函(2014)26号-16] （2014）
2016年度广东省大学生创新创业训练计划资助项目(201610576028) （201610576028）
