金黄色葡萄球菌模板制备的比较研究

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[中图分类号]Q939.1;TS 207.4[文献标志码]A[文章编号]1005-0310（2025）03-0045-08

Abstract：The present study displayed five diferent 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 10υ to 105 was successfully detected by conventional PCR，while template DNA extractedusing the commercial kits extraction methodand instrument automatic extraction at a 10∘ 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.

Keywords ： staphylococcus aureus ; PCR ;fluorescence quantitative PCR ; template DNA

金黄色葡萄球菌是一种典型的革兰氏阳性球型细菌，可以大量分布于空气、水、人和动物的排泄物中，富含蛋白质的食品如蛋奶类、肉类容易受到污染。（剩余9435字）