文献类型：期刊文献

中文题名：金黄色葡萄球菌模板制备的比较研究

英文题名：A Comparative Study of the DNA Template Preparation for Staphylococcus Aureus Detection via PCR

作者：商迎辉[1];李乾伟[1];黄汉昌[1];劳凤学[2]

第一作者：商迎辉

机构：[1]北京联合大学生物化学工程学院,北京100023;[2]北京联合大学应用科技学院,北京100012

第一机构：北京联合大学生物化学工程学院

年份：2025

卷号：39

期号：3

起止页码：45-52

中文期刊名：北京联合大学学报

外文期刊名：Journal of Beijing Union University

语种：中文

中文关键词：金黄色葡萄球菌;PCR;荧光定量PCR;模板DNA

外文关键词：staphylococcus aureus;PCR;fluorescence quantitative PCR;template DNA

摘要：通过实验系统比较了煮沸裂解法、SDS裂解法、改良SDS裂解法、商品化试剂盒提取法以及仪器自动提取法5种模板DNA制备方法在金黄色葡萄球菌PCR检验过程中的适用性。采用上述方法从金黄色葡萄球菌培养物中提取模板DNA并进行10倍系列稀释,应用各稀释度模板DNA进行常规PCR和荧光定量PCR检测。研究发现,SDS裂解法和改良SDS裂解法制备的模板DNA未经稀释时,在常规PCR和荧光定量PCR检测中均不能检出。煮沸裂解法制备的10^(0)~10^(4)稀释度的模板DNA使用常规PCR均被检出,而商品化试剂盒提取法以及仪器自动提取法制备的10^(4)稀释度的模板DNA使用常规PCR均未被检出。煮沸裂解法制备的10^(0)~10^(5)稀释度的模板DNA使用荧光定量PCR均被检出,其他4种方法制备的10^(5)稀释度的模板DNA使用荧光定量PCR均未被检出。煮沸裂解法制备的模板在PCR检验过程中的适用性好、操作简便、实验室要求低,是金黄色葡萄球菌PCR检验过程中模板制备的最优方法。
The present study displayed five different methods of template DNA extraction from Staphylococcus aureus cultures,including Simple boiling pyrolysis,SDS cracking,SDS combined with heat cracking,commercial kits extraction method and instrument automatic extraction.By using the conventional PCR and fluorescence quantitative PCR,the effects of the DNA template prepared by these methods for PCR test are compared.The study revealed that template DNA extracted using the SDS cracking and SDS combined with heat cracking methods was undetectable in both conventional PCR and fluorescence quantitative PCR without prior dilution.Template DNA extracted by simple boiling pyrolysis at dilutions ranging from 100 to 105 was successfully detected by conventional PCR,while template DNA extracted using the commercial kits extraction method and instrument automatic extraction at a 105 dilution was undetected.In fluorescence quantitative PCR,template DNA obtained via the boiling pyrolysis at dilutions from 100 to 105 was consistently detected,while DNA at a 105 dilution prepared by the other four methods mentioned above was undetected.Given its high suitability for PCR-based detection,simplicity,and minimal laboratory requirements,the boiling pyrolysis method was identified as the optimal approach for template DNA preparation in Staphylococcus aureus PCR assays.