环介导等温扩增技术结合横向流动试纸条检测猪链球菌2型的应用研究

发布时间：2018-01-19 02:02

本文关键词： 猪链球菌型 cpsJ基因环介导等温扩增横向流动试纸条　出处：《中国人兽共患病学报》2016年12期 　论文类型：期刊论文

【摘要】：目的建立一种猪链球菌2型的快速检测方法。方法以猪链球菌2型cps2J基因序列为检测靶标设计6条环介导等温扩增(LAMP)特异性引物,其中2条促环引物分别标记异硫氰酸荧光素(FITC)和地高辛(Dig),用横向流动试纸条(LFD)检测扩增产物,建立猪链球菌2型LAMP-LFD检测方法。结果所建立的检测方法能够特异性的检测出猪链球菌2型,与试验对照菌株不存在交叉反应,其最低检测限为76cfu/mL,在对100份样品的检测中,LAMP-LFD方法检测出14份阳性样品,培养法检测出12份阳性样品,LAMP-LFD方法检测阳性率略高于病原培养法,两者间的P0.05无统计学差异。结论所建立LAMP-LFD检测方法具有特异性好、敏感性高、操作简单、快速等特点,适合于动物食品中猪链球菌2型的检测。
[Abstract]:Objective to establish a rapid detection method for Streptococcus suis type 2. Methods six specific primers for isothermal amplification of streptococcus suis type 2 were designed using cps2J gene sequence of Streptococcus suis type 2 as the detection target. Two ring stimulating primers were labeled with fluorescein isothiocyanate (FITC) and digoxigenin (Digg) respectively, and the amplification products were detected by LFDs. The detection method of Streptococcus suis type 2 LAMP-LFD was established. Results the method was able to detect Streptococcus suis type 2 specifically, and there was no cross reaction with the control strain. The lowest detection limit was 76cfu-mL. in 100 samples, 14 positive samples were detected by LAMP-LFD and 12 positive samples were detected by culture method. The positive rate of LAMP-LFD method is slightly higher than that of pathogen culture method, there is no statistical difference between them (P0.05). Conclusion the established LAMP-LFD detection method has good specificity and high sensitivity. Simple and rapid operation, suitable for the detection of Streptococcus suis type 2 in animal food.
【作者单位】：天津市畜牧兽医研究所;中国兽医药品监察所;
【基金】：国家高技术研究发展计划资助项目(No.2012AA101605) 天津市农业科技成果转化与推广项目(No.201301030) 天津市农业科学院基金项目(No.16003)联合资助~~
【分类号】：R440
【正文快照】： (1.Tianjin Veterinary Research Insititution,Tianjin300384,China;2.China Institute of Veterinary Drug Control,Beijing100081,China)猪链球菌病(swine streptococcosis)是由C、D、F及L群链球菌引起的猪的多种疾病的总称,急性型常表现为出血性败血症和脑炎,慢性型以关节

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