利用LIVE/DEAD试剂盒染色法,流式细胞仪检测法,以及RT-PCR方法检测了经低浓度冰醋酸作用的鸡源大肠杆菌活细胞,并对其进行了复苏。结果表明,当可培养菌数降为零时,LIVE/DEAD试剂盒染色法及流式细胞仪检测法均能检测出较高数量的活菌数;RT-PCR法也能检测到活细胞信号分子-mRNA,并扩增出“活的非可培养状态”(VBNC)细菌的cDNA片段。这些均显示细菌已进入VBNC,且在吐温80作用下又恢复为可培养状态,从而证实大肠杆菌具有“VBNC”状态。
Viable but nonculturable state(VBNC) induced by low dimensional glacial acetic acid in avianEscherichia was detected by UVE/DEAD kit, flow cytometry and RT-PCR. And some resuscitation experiments were done for VBNC avian Escherichia. Results indicated that the titer of CFU declined to undetectable levels,while more viable avian Escherichia were determined by UVE/DEAD kit and flow cytometry count methods. And live cell signaling molecular-mRNA was also detected by RT-PCR, and more cDNA fragments were amplificated. These results indicated that avian Escherichia had entered into the viable but nonculturable state. And avian Escherichia cells would be recoverable from a viable but nonculturable state by Tween 80. Above mentioned research provides the foundation for futher research VBNC bacteria genome and diffrence gene between VBNC and normal state.