单增李斯特菌非可培养状态的诱导与PMA-qPCR检测

Induction and PMA-qPCR detection of Listeria monocytogenes in viable but non-culturable state

在9%(w/v)NaCl条件下,通过温度、pH两种因素诱导一株单增李斯特菌进入"活的非可培养"(viable but non-cuturable,VBNC)状态。通过LIVE/DEAD Baclight活菌检测试剂盒检测细菌总数与活菌数,比较单增李斯特菌状态及数量的差异与变化。结果显示,在2℃、pH6.0条件下单增李斯特菌活菌全部进入VBNC状态。用PMA-qPCR对经诱导的四份菌液进行检测,结果表明该方法能快速、准确地测出死菌背景下的活的单增李斯特菌。

Under the condition of 9% （w/v）NaCI, the induction of ＂viable but non culturable （VBNC）Listeria monocytogenes was undertaken by investigating two factors, temperature and pH. In order to compare the changes of bacteria state and numbers caused by different inducing conditions, the LIVE/DEAD ~ BacLightTM Bacterial Viability Kit was used for enumeration of total cells and viable cells. The results showed that all viable Listeria monocytogenes were induced into the VBNC state at 2~C, pH6.0.PMA-qPCR was proposed to detect the Listeria monocytogenes experienced different inductions.It was indicated that PMA-qPCR was an effective method to detect viable Listeria monocytogenes even if undering the dead cells background.