细菌生理特性快速检测试剂盒研制

A kit for rapid detection of bacterial physiological characters

选取各代谢途径中利用某些特定碳源,如蔗糖、葡萄糖、甘露醇等95种碳源作为测试碳源,分别加入96孔酶标板中。选用指示剂噻唑兰(MTT)来鉴定细菌利用特征碳源情况。同时选取不同种类和不同浓度的抗生素,不同起始pH、不同NaCl浓度加入96孔酶标板中,测定细胞是否生长,可以快速地获得该菌的抗生素耐受性谱。试验还从指示剂浓度、培养基的固化、接菌浓度、观察时间等方面对试剂盒进行优化,得到一种最佳的试剂盒鉴定系统BiobiqA(碳源利用谱)和BiobiqB(生理抗性谱)。用模式菌株对试剂盒进行测试和验证,结果表明,试剂盒具有操作简便、结果准确、节省成本、节约时间等优点,可以进行细菌生理特性的快速检测。

To quickly detect bacterial physiological characters, the authors developed a kit to test the utilization of various carbon sources by bacteria and their responses to medium pH and antibiotics. In the kit, 95 kinds of carbon sources were utilized for target bacteria, and the indicator MTF was used to test the utilization of specific carbon sources by the target bacteria. The growth of target bacteria at a series of medium initial pH and NaC1 concentration and the sensitivities of the bacteria to various kinds and different concentrations of antibiotics were also tested. For result observation, an optimization was made on the MTT concentration, medium solidification, bacterial inoculation amount, and observation duration. Accordingly, a kit with BiobiqA （C-source uti- lizing spectra） and BiobiqB （antimicrobial sensitivity pattern） was developed. The test and validation with model bacteria （Enterobacter cloacae ATCC13047T and Rhizobium NZP2213T） showed that our newly developed kit had the advantages of broad spectrum, easy operation, high accuracy, costsaving, and time-saving, being able to rapidly detect the physiological characters of bacteria