酶标仪分光光度法在微生物耐性研究上的应用

The Application of Microplate Reader Spectrophotometry in Microbial Tolerance Analysis

微生物在环境中的耐性是该微生物在相应环境发挥作用的重要基础。为了获得一种用于微生物耐性分析的快速简便方法,传统平板分离法和酶标仪分光光度法被用于评估其在细菌和链霉菌紫外耐受水平检测中的差异,并分析了酶标仪分光光度法在微生物其他耐性水平检测中的适用性。结果显示,两种检测方法均能体现细菌和链霉菌对紫外线的耐受水平,前者经过稀释、涂布、培养、菌落计数,获得的是菌株在紫外线照射后的存活浓度,而后者经过接种96孔培养板、培养、吸光度检测,获得的是菌株经紫外线照射后的生长曲线,并从生长曲线获知菌株的生长速率、增殖能力等信息。此外,酶标仪分光光度法同样适用于细菌对pH和盐的耐受水平分析,对于链霉菌耐受性分析有一定的适用性。酶标仪法除了能获得与平板分离法相似的耐性水平检测外,还能获得菌株在不同耐性水平上的增殖潜力,且在操作上比平板分离法省时、省力,可用于微生物耐性分析、高通量筛选等研究工作。

Microbial tolerance in environment is an important base in playing its role in the corresponding environment.In order to obtain a rapid and handy method used in microbial tolerance analysis,traditional plate separation and microplate reader spectrophotometry(MRS)were adopted to appraise the differences in the determination of UV tolerance levels of bacteria and streptomyces,and analyze the applicability of the MRS in the determination of other tolerance level of microbes.The results showed that both of the two detection methods can reflect the tolerance level of bacteria and streptomyces to UV.The concentration of the survival strain after UV irradiation could be obtained by the former method through diluting,spreading,culturing,and colony counting,while the growth curve of the survival strain after UV irradiation could be obtained by the latter method through inoculating in 96 hole culture plate,cultivation,absorbance detection,then the growth rate,proliferation ability and other information of the strain could also be obtained from the growth curve.In addition,the MRS method was also suitable for the analysis of bacterial pH and salt tolerance,and had certain applicability for the analysis of streptomyces tolerance.As compared with the plate separation method,not only the similar tolerance level,but also the proliferation potential of the strains at different tolerance levels could be obtained,and more time and effort can be saved by the microplate reader method,which is suitable for the research work on microbial tolerance analysis and high-throughput screening,etc.