优化血小板制品中细菌基因组抽提方法

Optimization of Methods for Extracting Bacterial Genomic DNA in Platelet Products

目的:建立裂解液加热煮沸法抽提细菌基因组DNA。方法:选取大肠埃希菌、金葡菌分别应用五种不同方法抽提细菌基因组,Tagman荧光定量PCR检测抽提产物。并在两种细菌中验证chelex-100抽提方法的灵敏度。结果:对相同浓度的大肠埃希菌、金葡菌样品进行抽提时,chelex-100加热煮沸法抽提所得样品在后续荧光定量PCR检测中CT值均为最小。应用该方法对两种细菌进行抽提鉴定时,灵敏度可达到每毫升个位菌数。结论:chelex-100裂解液加热煮沸法抽提细菌基因组DNA操作简便、省时、经济,为血液标本细菌基因检测在临床上的大规模应用奠定基础。

Objective： To establish new methods to extract bacterial genomic DNA. Methods： The same concentration E. coli and S. aureus were prepared with five methods for isolation of genomic DNA. Then, the genomic DNA was detected by using Tagman Real - Time PCR. And confirm the sensitivity of chelex - 100 - boiling in these two bacteria. Results：Among of these methods, the values of CT from chelex - 100 - boiling method were the least when extract genomic DNA from the same concentration E. coli and S. aureus. Single E. coli and S. aureus could be detected using the method. Conclusion： The chelex- 100- boiling method to extract genomic DNA from bacteria is convenient, less time and consuming, and may be applied to large scale of clinical detection of bacteria.