摘要
目的建立环介导等温扩增技术(LAMP)快速检测霍乱弧菌。方法针对霍乱弧菌外膜蛋白的ompW基因设计特异引物,优化反应条件,建立霍乱弧菌的LAMP检测方法。通过对47株细菌及模拟污染现场,检测该方法的灵敏度、特异度和实用性。结果活菌计数方法表明建立的LAMP方法检测霍乱弧菌的灵敏度为1.6×102cfu/ml,在人工污染霍乱弧菌的粪便及海水标本中检测的敏感度为1.6×103cfu/ml,在牛奶标本中敏感度为1.6×104cfu/ml。检测21株霍乱弧菌均为阳性,26株非霍乱菌则全部阴性,特异度为100%。结论建立的LAMP方法检测霍乱弧菌灵敏度、特异度高,可用于霍乱现场监测和流行病学调查。
Objective To establish a loop-mediated isothermal amplification （LAMP） method for rapid diagnosis of Vibrio cholerae. Method Based on the ompW nucleic sequence ofVibrio cholerae, a pair of primers was designed for LAMP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of LAMP were tested using 47 baterial strains and simulated contaminated sites. Results The results of viable bacterium count showed that LAMP was capable of detecting Vibrio cholerae at a level as low as 1.6×10^2 cfu/ml. The minimal detectable concentration was 1.6×10^3 cfu/ml for simulated contaminated samples such as feces and seawater, and 1.6×10^4 cfu/ml for contaminated milk. All the 21 strains of Vibrio cholerae yielded positive results in LAMP, and the 26 strains of other bacteria all showed negative results, with a detection specificity of 100%. Conclusion The established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第10期2059-2063,共5页
Journal of Southern Medical University
关键词
霍乱弧菌
环介导等温扩增技术
快速检测
Vibrio cholerae
loop-mediated isothermal amplification
rapid diagnosis
