中国兽医科学2024，Vol.54Issue(8)：1043-1049,7.DOI:10.16656/j.issn.1673-4696.2024.0131
基于布鲁菌bp26基因LAMP检测方法的建立及优化
Establishment and optimization of LAMP detection system in Brucella based on bp26 gene
摘要
Abstract
Brucellosis seriously affects the development of animal husbandry and public health safety.Timely and accurate detection of brucellosis is the key to the prevention and control of the disease.The LAMP(loop-mediated isothermal amplification)primers were designed based on the Brucella bp26 gene,to establish and optimize the Brucella LAMP assay in this study.To obtain the best reaction conditions,magnesium ion concentration,LAMP primer concentration,reaction equipment,and other reac-tion conditions were optimized,and the sensitivity and specificity of the LAMP assay were evaluated,respectively.The optimal LAMP assay was established when the magnesium ion concentration was 8 mmol/L and the ratio of internal and external primers was 6∶1.There was no difference in the metal bath,ther-mostatic water bath,and oven.This method could only detect Brucella and does not react with Es-cherichia coli,Streptococcus pneumoniae,and Staphylococcus aureus etc,indicating that the estab-lished LAMP assay had good specificity.Under the optimal reaction conditions,the detection sensitivi-ty was 10 pg/μL.For obtaining a good visual detection effect,the concentration of different concen-trations of manganese ions was optimized in the reaction system,and the results showed that 0.1 mmol/L manganese ions could establish the best visual effect.The Brucella LAMP assay is specific,sensitive,and visual,providing technical support for rapid clinical detection of Brucella.关键词
布鲁菌/环介导等温扩增/bp26/可视化检测Key words
Brucella/LAMP/bp26/visibility test分类
农业科技引用本文复制引用
马世辉,贾沅铮,王福成,连丽燕,蒋蔚,陈兆国,韩先干,陈伟..基于布鲁菌bp26基因LAMP检测方法的建立及优化[J].中国兽医科学,2024,54(8):1043-1049,7.基金项目
上海市科技创新行动计划"农业科技领域项目"(0301001999379,21N31901000) （0301001999379,21N31901000）
中央级公益性科研院所基本科研业务费专项资金项目(2023JB03) （2023JB03）
