摘要
目的：比较荧光定量PCR法和培养法在结核分枝杆菌定量上是否有差异及探讨应用DNA拷贝浓度替代菌液浓度的可能性。方法：将6种不同浓度的H37Rv标准菌株悬菌液各分为两组：PCR法组和培养法组，并在同一个721型可见光分光光度仪上测定其OD600值。然后，采用荧光定量PCR法，建立结核分枝杆菌DNA拷贝浓度OD标准曲线，同时对同一标本进行培养并行菌落计数，比较PCR方法和培养法对结核分枝杆菌定量结果是否一致。结果：两种方法对结核分枝杆菌定量无显著性差异（P〉0．05）。OD值与DNA拷贝浓度和悬菌液浓度均呈正相关，相关系数分别为1，P〈0．01。其回归方程PCR法为：C（H37 Rv）=-0．0189＋6．9910D，培养法为：C（H37Rv）=-0．0201＋6．7570D，OD范围0．1～0．9。结论：荧光定量PCR法和培养法所得结果基本一致，是一种结核分枝杆菌定量新的方法，可替代菌液浓度法应用于动物实验。
Objective： To compare the fix quantity of mycobacterium tuberculosis by the fluorescent quantitative PCR method and the culture method and to explore the probability of replacing concentration of DNA for concentration of bacterium. Methods： Six standard vaccine emvlsions of H37Rv of different conentration were divided into two groups ： the PCR group and the culture group and their OD600, value under the same spectrophotometer was measured . Then, the fluorescent quantitative PCR method was used to establish a standard OD curve of mycobacterium tuberculosis- concentration of DNA. Meanwhile the same sample was cultured to count the bacterium to compare the fix quantifying results of mycobacterium tuberculosis by the fluorescent quantitative PCR method and the culture method. Results ： There is no significant difference in the quantity of mycobacterium tuberculosis by the above two methods （P〈0.05）. The value of OD was positively related to concentration of DNA and bacterium respectively and both of the correlation coefficients were 1,P〈0.01. The regression equation of the PCR group was ;C （H37Rv） = - 0. 0189 ＋ 6. 991OD ;the regression equation of the culture group was ：C （H37Rv） = - 0. 0201 ＋ 6. 757OD. Conclusion.The results of the PCR method and the culture method are the same to a large degree： so it is a new fix quantifying method to replace concentration of bacterium for the animal experiments.
出处
《陕西医学杂志》
CAS
北大核心
2006年第9期1171-1172,1181,共3页
Shaanxi Medical Journal
基金
西安市科技攻关项目(GG05144)
关键词
分枝杆菌
结核/诊断
聚合酶链反应
荧光光度测定
细胞
培养的
Mycobacterium, tuberculosis/ diagnosis Polymerase chain reaction Fluorophotometry Cell, cultured
