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目的探讨用96孔酶标板法检测家蝇血淋巴黑化反应的可行性。方法用大肠埃希菌、金黄色葡萄球菌、藤黄微球菌单菌和大肠埃希菌与金黄色葡萄球菌混合菌分别体外刺激家蝇3龄幼虫的血淋巴,通过96孔酶标板法分析菌刺引起的家蝇幼虫血淋巴酚氧化酶活性的变化,进一步探讨菌刺与家蝇黑化反应的关系。结果相比正常对照的起始A490值,1~3号样品的起始A490值随加菌量的增多依次递增,其增幅分别为大肠埃希菌1.33~1.38倍,金黄色葡萄球菌1.30~2.40倍,藤黄微球菌2.60~3.00倍,大肠埃希菌与金黄色葡萄球菌混合菌1.40~3.80倍。显然,菌刺可使家蝇血淋巴酚氧化酶活性增高,菌刺浓度越大,黑化反应越强,单菌刺激的效果依次为藤黄微球菌>金黄色葡萄球菌>大肠埃希菌,大肠埃希菌与金黄色葡萄球菌混合菌刺激的效果要比单菌好。结论用96孔酶标板法检测菌刺家蝇血淋巴黑化反应的变化可行。
Objective To explore the feasibility of using 96-well microtiter plate assay to detect hemolymph of housefly. Methods The hemolymph of housefly 3rd instar larvae was stimulated in vitro with Escherichia coli, Staphylococcus aureus, Micrococcus luteus, Escherichia coli and Staphylococcus aureus respectively, and then analyzed by 96- The changes of hemolymph phenoloxidase activity in housefly larvae induced by the stab were further discussed, and the relationship between the stab and the blackfly reaction in housefly was further explored. Results Compared with the initial A490 value of the normal control, the initial A490 values of samples 1-3 increased with the increase of the amount of bacteria, which were 1.33-1.38 times of those of Escherichia coli and 1.30-2.40 of Staphylococcus aureus Times, Micrococcus luteus 2.60 ~ 3.00 times, Escherichia coli and Staphylococcus aureus mixed bacteria 1.40 ~ 3.80 times. Obviously, the activity of hemolymph phenoloxidase in Housefly increased obviously with the increase of the concentration of stab, and the stronger of blackening reaction. The order of single bacteria stimulation was Micrococcus luteus> Staphylococcus aureus> Escherichia coli, Escherichia coli and Staphylococcus aureus mixed bacteria to stimulate the effect is better than single bacteria. Conclusion It is feasible to detect the change of hemolytic reaction of Musca domestica with 96-well plate.