Quantitative PCR coupled with sodium dodecyl sulfate and propidium monoazide for detection of viable Staphylococcus aureus in milk

文献类型： 外文期刊

第一作者： Dong, Lei

作者： Dong, Lei;Liu, Huimin;Meng, Lu;Xing, Mengru;Wang, Jiaqi;Zheng, Nan;Dong, Lei;Liu, Huimin;Meng, Lu;Xing, Mengru;Wang, Jiaqi;Zheng, Nan;Dong, Lei;Xing, Mengru;Liu, Huimin;Wang, Cheng;Chen, He

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关键词： propidium monoazide; sodium dodecyl sulfate; Staphylococcus aureus; quantitative polymerase chain reaction; internal amplification control

期刊名称：JOURNAL OF DAIRY SCIENCE （ 影响因子：4.034； 五年影响因子：4.354 ）

ISSN： 0022-0302

年卷期： 2018 年 101 卷 6 期

页码：

收录情况： SCI

摘要： Conventional quantitative PCR (qPCR) are unable to differentiate DNA of viable Staphylococcus aureus cells from dead ones. The aim of this study was to use sodium dodecyl sulfate (SDS) and propidium monoazide (PMA) coupled with lysostaphin to detect viable Staph. aureus. The cell suspensions were treated with SDS and PMA before DNA extraction. The SDS is an anionic surfactant, which can increase the permeability of dead cells to PMA without compromising the viability of live cells. The lysostaphin was applied to improve the effectiveness of DNA extraction. The reliability and specificity of this method were further determined by the detection of Staph. aureus in spiked milk. The results showed that there were significant differences between the SDS-PMA-qPCR and qPCR when a final concentration of 200 mu g/mL of lysostaphin was added in DNA extraction. The viable Staph. aureus could be effectively detected when SDS and PMA concentrations were 100 mu g/mL and 40 mu M, respectively. Compared with conventional qPCR, the SDS-PMA-qPCR assay coupled with lysostaphin was more specific and sensitive. Therefore, this method could accurately detect the number of viable Staph. aureus cells.

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