摘要
本研究建立了一种快速诊断动物大肠杆菌病的PCR检测方法,与传统的细菌分离培养、生化鉴定方法相比较有效缩短了检测时间,提高了检测效率,同时为大肠杆菌病的快速诊断和流行病学调查提供了有效的技术手段。通过对大肠埃希菌ECs1048基因序列分析,利用Premier5.0软件设计并合成了1对特异性引物,建立了检测大肠埃希菌ECs1048基因的PCR检测方法。通过反复试验确定本方法的最佳退火温度为56℃。灵敏性试验和特异性试验表明本方法能够检测出的最低菌悬液浓度为1.5×10^2 CFU/mL,并具有较高的特异性。稳定性与重复性试验表明本方法具有良好的稳定性。利用本方法对临床样品的检测结果与生化试验鉴定结果的符合率为100%。本研究建立的检测方法灵敏性高、特异性强、稳定性好,可应用于动物大肠杆菌病的诊断。
This study established a rapid diagnostic PCR method for colibacillosis in animals.Compared with traditional bacterial culture, biochemical identification method, the PCR can effectively reduce the test time,improve the detection efficiency.Also it provided the effective technical means for rapid diagnosis and epidemiological investigation of the disease.Based on ECs1048 gene sequence from Escherichia coli ,a pair of specific primers were designed and synthesized by premier 5.0 software,and the PCR detection method for ECs1048 gene was established.The optimum annealing temperature of the method was 56℃ by repeated experiments.Sensitivity and specificity tests showed that the method can detect the minimum suspension concentration 1.5 × 10^2 CFU/mL, and has a high specificity.Stability and repeatability tests showed that the method has good stability.At the same time,the method was applied to detect the clinical samples,and the coincidence rate of the test results and the biochemical test results was 100%.Thus,The detection method established in this study has high sensitivity,strong specificity and good stability.It can be used in the labo- ratory diagnosis of colibacillosis in animals.
出处
《动物医学进展》
北大核心
2016年第8期11-14,共4页
Progress In Veterinary Medicine
基金
辽宁省自然科学基金项目(201402573)
辽宁省农业攻关及产业化项目(2015202013)
关键词
大肠埃希菌
聚合酶链反应
建立
Escherichia coli
polymerase chain reaction
establishment
