一种细菌DNA提取方法的正交设计优化  

Optimizing a method of extracting bacteria DNA by orthogonal design

作　　者：张锦伟[1] 陈苏红[2] 刘琪琦[2] 张宏刚[3] 王升启[2] 姜华[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029 [2]军事医学科学院放射与辐射医学研究所,北京100850 [3]山西医科大学微生物与免疫教研室,山西太原030001

出　　处：《军事医学科学院院刊》2010年第6期583-585,588,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家传染病重大专项资助项目(2008ZX10004-02)

摘　　要：目的通过正交实验优化异硫氰酸胍-硅胶提取细菌DNA的方法,以缩短其提取时间,进而缩短细菌特别是炭疽芽孢杆菌等恐怖细菌的核酸分子检测时间。方法以基因芯片的杂交信号值为指标,以异硫氰酸胍-硅胶提取DNA过程中的煮沸时间、室温放置时间、乙醇洗涤次数为考察因素通过正交实验法优化异硫氰酸胍-硅胶提取细菌DNA的条件。结果优化后的因素水平为煮沸10 min,室温放置15 min,乙醇洗涤2次。结论实验证明,优化后的提取时间比原来缩短了25 min,且信号值比原方法更高。Objective To optimize guanidinium isothiocyanate-based-silicon dioxide extraction of bacteria DNA in order to shorten the time of extraction,especially the time taker to detect nucleic acid of bio-terrorism bacteria,such as Bacillus anthracis.Methods The time of guanidinium isothiocyanate-based-silicon dioxide extraction of bacteria DNA could be optimized by orthogonal experiments with the hybridized value as the index and with boiling time of guanidinium isothiocyanate-based-silicon dioxide extraction of bacteria DNA,duration of storage at room temperature,and times of washing by ethanol solution as considerations.Results The optimized condition was ten minutes of boiling,fifteen minutes of storage at room temperature and twice washing by ethanol solution.Conclusion This experiment can confirm that the optimized time of extraction is cut down by twenty minutes and signal value is higher compared with the original method.

关 键 词：正交实验 基因芯片 实时荧光定量PCR 寡核苷酸序列分析 炭疽 

分 类 号：R440[医药卫生—诊断学]