金黄色葡萄球菌活的非可培养状态的诱导和检出

Analysis of Viable But Non-culturable State of Staphylococcus aureus

目的研究金黄色葡萄球菌"活的非可培养状态"(viable but non-culturable state,VBNC)的诱导和检出方法。方法将107cfu/ml的金黄色葡萄球菌ATCC13565分别采用4℃冷藏、-20℃冷冻及添加0.01～0.05mmol/L铜离子的方法进行诱导,并对VBNC菌体进行了检测;采用逐步升温和化学促进法对VBNC菌体进行复苏试验。结果-20℃低温冷冻诱导72h或经0.015mmol/LCu2+诱导4d均可诱导菌体进入VBNC;在培养基中添加0.5%吐温20或1%过氧化氢酶培养24h均可使处于VBNC的菌体实现复苏。结论金黄色葡萄球菌可以通过诱导进入VBNC;而经过复苏后的金黄色葡萄球菌与正常菌体在通用的检验培养基中菌落形态和生理生化反应均一致。

Objective To study the induction and detection methods of Staphylococcus aureus viable but non-cuhurable state （VBNC）. Methods Staphylococcus aureus （ATCC13565） of 10^7 cfu/ml was induced by low temperature at 4 ℃, freezing at -20 ℃ and adding copper ions from 0.01 mmol/L to 0.05 mmol/L, Staphylococcus aureus was tested, and explored the resuscitation conditions by stepwise heating-heating and chemokinesis. Results Frozen 72 h at the temperature of -20℃ or added 0.015 mmol/L Cu^2＋ for 4 days of culture could induce staphylococcus aureus into the state of VBNC. Adding 0.5% tween-20 or 1% catalase for 24 h of culture could make the strain in VBNC achieve recovery. Conclusion Staphylococcus aureus can be induced into the VBNC and the recovered Staphylococcus aureus is the same as the normal bacteria in colony morphology and physiological and biochemical response in the common test medium.