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目的:基于明亮发光杆菌502和青海弧菌Q67的Microtox微毒测试体系比较研究,明确两者适宜的反应条件。方法:1考察明亮发光杆菌502和青海弧菌Q67在15℃±1℃、20℃±1℃、25℃±1℃下的相对发光情况;2采用Plackett-Burman法优化设计影响明亮发光杆菌502和青海弧菌Q67发光的因素;3采用Plackett-Burman法优化得到的反应体系,考察不同p H复苏液对上述两种细菌相对发光强度的影响。结果:1明亮发光杆菌502的相对发光强度随温度的升高而增强,而青海弧菌Q67的相对发光强度随温度的升高先增强后减弱。2Plackett-Burman法得到最优发光条件为:明亮发光杆菌502冻干粉平衡10min后加入复苏液2ml,混匀15min,取100μl菌液测试初始发光度,随后立即加入1ml复苏液或待测样品,反应15min后再次测试相应发光强度;青海弧菌Q67冻干粉平衡10min后加入复苏液2ml,混匀10min,取100μl菌液测试初始发光度,随后立即加入2ml复苏液或待测样品,反应10min后再次测试相应发光强度。3复苏液p H 3.6-p H 3.8时对明亮发光杆菌502发光的影响由抑制转为增强;复苏液p H 4.5-p H 5.0时对青海弧菌Q67发光强度的影响<±15%。结论:与明亮发光杆菌502比较,青海弧菌Q67具有更宽的p H耐受范围并且在检测时无需调节待测样品渗透压。
OBJECTIVE: To determine the appropriate reaction conditions of Microtox microtitre assay system based on the comparison of Luminescent bacterium 502 and Q67. Method 1: The relative luminescence of photobacteriumsbacterium luminarum 502 and Q67 of Qhaihai at 15 ℃ ± 1 ℃, 20 ℃ ± 1 ℃ and 25 ℃ ± 1 ℃ were investigated.2. And Q67 of Qinghai seaweed.3. The reaction system optimized by Plackett-Burman method was used to investigate the effect of different p H resuscitation solutions on the relative luminescence intensity of the two bacteria. Results: 1 The relative luminescence intensity of Brilliant Brightobacteria 502 increased with the increase of temperature, while the relative luminescence intensity of Q67 increased firstly and then decreased with the increase of temperature. 2Plackett-Burman law to obtain the optimal luminescent conditions: Liang Luminescent bacterium 502 freeze-dried powder balance 10min after adding the recovery solution 2ml, mix 15min, take 100μl bacterial liquid test initial luminosity, then immediately add 1ml resuscitation solution or test sample, After reaction for 15min, the corresponding luminescence intensity was tested again; Vibrio Q67 was added to the reconstituted solution 2ml after mixing for 10min, and mixed for 10min. The initial luminosity was tested with 100μl bacterial liquid, and immediately afterwards, 2ml resuscitation solution or test sample was added and reacted for 10min After the test again the corresponding luminous intensity. 3 resuscitation solution p H 3.6-p H 3.8 on the luminescence of photobacterium photobacterium photolyticus 502 was changed from inhibition to enhancement; the effect of resuscitation solution p H 4.5-p H 5.0 on the luminescence intensity of Q67 was less than ± 15%. Conclusion: Compared with the bright light-emitting bacteria 502, Vibrio Qinghai Q67 has a wider range of p H tolerance and no need to adjust the osmotic pressure of the sample to be tested.