微生物ATP提取方法及其在微波灭菌效果快速评价中的应用研究

Optimization and application of ATP Bioluminescence System in the Rapid Evaluation of Microwave Sterilization

DOI：

中文关键词: 生物发光 苯扎溴铵 微波灭菌 快速检测

英文关键词: bioluminescence benzalkonium bromide(BAB) microwave sterilization rapid detection

基金项目:国家863计划项目(2008AA10Z419);国家自然科学基金项目(20805019);江南大学科学基金项目

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中文摘要:

本文对ATP发光技术中微生物ATP提取方法进行了优化研究,通过对不同提取剂提取效率的比较,发现苯扎溴铵提取效果好,并且无需稀释,能真实体现微生物细胞内的ATP水平。通过对苯扎溴铵提取时间和提取浓度的优化,结果显示采用体积分数为0.05%苯扎溴铵溶液与菌液混合3~4 min,即可将细胞内的ATP最大限度地释放出来。将该提取方法与ATP生物发光反应体系偶联,实现了对微波灭菌效果的快速评价,在短时间内即可得到可靠结果。试验比较了不同微波功率和微波时间的灭菌效果,实验结果表明,大肠杆菌ATCC25922在用2450 MHz 650w微波照射60 s时可被完全杀灭,而金黄色葡萄球菌ATCC25923则需70 s才能被杀灭。当用2450 MHz微波同样处理60 s时,650 w即可杀灭大肠杆菌ATCC25922,而金黄色葡萄球菌ATCC25923则需700 w才可被完全杀灭。

英文摘要:

In this manuscript,the extraction method of microorganism ATP used in bioluminescence techniques were studied.According to compare the extract results of different extractants,it was found that that benzalkonium bromide(BAB) is the best extractant to extract all of ATP microorganism cells.The extraction conditions including the extractant concentration and the extaction time were determined and listed as follows: 0.05 % BAB mixing with bacteria cells for 3-4 min.The finding was applied to rapidly evaluate the effect of microwave sterilization coupling with bioluminescent technology and the reliable results can be obtained within a short time.According to the sterilization effect of different microwave power and sterilization time,we found that E.Coli ATCC25922 can be completely exterminated irrediated by 2 450 MHz 650 w microwave for 60 s but 70 s for Staphylococcus aureus ATCC25923.When treated with 2450 MHz microwave all for 60 s,the power for compelte extermination of E.Coli ATCC25922 is 650 w but 700 s for Staphylococcus aureus ATC25923.

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