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 μg/mL of lysostaphin was added in DNA extraction. The viable Staph. aureus could be effectively detected when SDS and PMA concentrations were 100 µg/mL and 40 μM, 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.

常规定量PCR（qPCR）不能区分存活的金黄色葡萄球菌细胞和死亡的DNA 。这项研究的目的是使用十二烷基硫酸钠（SDS）和单叠氮化丙锭（PMA）结合溶葡萄球菌素来检测可行的葡萄球菌。金黄色的。DNA提取前，将细胞悬液用SDS和PMA处理。SDS是一种阴离子表面活性剂，可以增加死细胞对PMA的通透性，而不会损害活细胞的活力。溶葡萄球菌素被用于提高DNA提取的效率。通过金黄色葡萄球菌的检测进一步确定了该方法的可靠性和特异性。金黄色在加牛奶的牛奶中。结果表明，当在DNA提取中添加最终浓度为200μg/ mL的溶葡萄球菌素时，SDS-PMA-qPCR和qPCR之间存在显着差异。可行的葡萄球菌。金黄色葡萄球菌可能当SDS和PMA浓度为100微克/毫升和40μ有效地检测中号分别。与传统的qPCR相比，SDS-PMA-qPCR分析法与溶葡萄球菌素联用更为特异性和灵敏性。因此，该方法可以准确地检测出可行的葡萄球菌数。金黄色细胞。