A novel loop-mediated isothermal amplification (LAMP) method for detection of Fusobacterium necrophorum from footrot

作者：Sun, D. B.; Wu, R.; He, X. J.; Zheng, J. S.; Wang, J. F.; Zhu, D. B.; Zhao, X. Y.; Guo, T.; Sun, B.; Fan, C. L.; Guo, D. H.^*

来源：African Journal of Microbiology Research, 2010, 4(23): 2617-2621.

• Fusobacterium necrophorum
• footrot
• loop-mediated isothermal amplification (LAMP)

A loop-mediated isothermal amplification method (Fn-LAMP) was developed for detection of Fusobacterium necrophorum from footrot using a set of four specific primers designed from the lktA gene of Fusobacterium necrophorum. The Fn-LAMP method, performed for 45 min at 61 degrees C, was capable of detecting 1 cfu/mL of F. necrophorum and was 10 times more sensitive than conventional PCR method. The Fn-LAMP had no cross-reaction with enterotoxigenic Escherichia coli, Salmonella, Pasteurella multocida, Mycoplasma, Campylobacter fetus from cattle. Detection of clinical samples indicated relative sensitivity and relative specificity of Fn-LAMP to PCR was 100 and 87.5%, respectively. The percentage of observed agreement was 92.0% between Fn-LAMP and PCR methods. Visual detection of the Fn-LAMP products showed negative and positive results could be clearly confirmed using fluorescent dyes as detection reagent.

• 出版日期2010-12-4
• 单位黑龙江八一农垦大学