本文選題：表皮葡萄球菌 + agrC��； 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]研究表皮葡萄球菌agrC特異結(jié)合多肽對聚氯乙烯材料表面細(xì)菌生物膜形成的作用,為合成agrC為靶點(diǎn)治療表皮葡萄球菌生物膜形成相關(guān)感染的特異性藥物奠定基礎(chǔ)。[方法]1.采用微量板半定量法確定agrC特異結(jié)合多肽的最佳抑菌濃度;2.采用微量板半定量法觀察agrC特異結(jié)合多肽對表皮葡萄球菌不同生長時期生物膜形成能力的影響;3.利用掃描電鏡觀察PVC材料表面細(xì)菌生物膜的表面結(jié)構(gòu);4.利用激光共聚焦顯微鏡觀察PVC材料表面細(xì)菌群落數(shù)量、細(xì)菌生物膜形成的厚度和細(xì)菌生物膜的三維結(jié)構(gòu)。[結(jié)果]1.微量板半定量法結(jié)果顯示表皮葡萄球菌agrC特異結(jié)合多肽的最佳抑菌濃度為 800μg/ml;2.微量板半定量法結(jié)果顯示當(dāng)agrC特異結(jié)合多肽濃度為800 μg/ml時,其在12h時對細(xì)菌生物膜的形成有明顯抑制作用;3.掃描電鏡觀察顯示表皮葡萄球菌ATCC35984在培養(yǎng)6h時,實驗組與對照組均可見散在小的細(xì)菌團(tuán)塊;在培養(yǎng)12h時,實驗組細(xì)菌團(tuán)塊明顯少于對照組;在18h時兩組均可見細(xì)菌團(tuán)塊相互連接成片狀、塔狀結(jié)構(gòu);在24h時兩組均可見成熟生物膜結(jié)構(gòu),結(jié)構(gòu)中可見大量無定形細(xì)胞外基質(zhì)填充;在30h時,兩組可見生物膜結(jié)構(gòu),但有部分呈分解狀。表皮葡萄球菌ATCC12228在培養(yǎng)6、12、18、24和30h時,實驗組和對照組細(xì)菌均未見生物膜形成。 ,4.激光共聚焦顯微鏡觀察顯示,實驗組加入agrC特異結(jié)合多肽與對照組加入無關(guān)肽,培養(yǎng)12h時觀察實驗組PVC材料表面細(xì)菌明顯少于對照組,且以死菌居多;而在6、18、24h兩組細(xì)菌量未見明顯差別,均以活菌居多。培養(yǎng)12h時觀察實驗組細(xì)菌生物膜厚度明顯少于對照組,其余時間點(diǎn)生物膜厚度未見明顯差別。[結(jié)論]1.表皮葡萄球菌與agrC特異結(jié)合多肽的抑菌強(qiáng)度之間存在劑量效應(yīng)關(guān)系;2. agrC特異結(jié)合多肽在生物膜形成的聚集階段對表皮葡萄球菌生物膜的形成有明顯抑制作用:agrC特異結(jié)合多肽能抑制表皮葡萄球菌生物膜的形成能力;agrC特異結(jié)合多肽可以阻礙表皮葡萄球菌進(jìn)一步聚集,使細(xì)菌團(tuán)塊疏松,抑制生物膜的形成;agrC特異結(jié)合多肽可以在細(xì)菌生物膜形成的聚集階段抑制細(xì)菌增殖,加速細(xì)菌死亡,影響細(xì)菌生物膜形成的厚度。3. agrC特異結(jié)合多肽可以為治療表皮葡萄球菌生物膜形成相關(guān)感染的特異性藥物奠定基礎(chǔ),它作為預(yù)防用藥可能更為有效,對成熟生物膜的抑制作用不明顯。
[Abstract]:[objective] to study the effect of agrC binding polypeptide of Staphylococcus epidermidis on bacterial biofilm formation on the surface of polyvinyl chloride (PVC), so as to lay a foundation for the synthesis of agrC as a target for the treatment of staphylococcus epidermidis biofilm formation related infection.[methods] 1.The optimum inhibitory concentration of agrC specific binding peptide was determined by microplate semi-quantitative method.The effect of agrC specific binding peptide on biofilm formation of Staphylococcus epidermidis at different growth stages was observed by microplate semi-quantitative method.The surface structure of bacterial biofilm on the surface of PVC was observed by scanning electron microscope (SEM).The number of bacterial community, the thickness of bacterial biofilm formation and the three-dimensional structure of bacterial biofilm on the surface of PVC were observed by laser confocal microscope.[result] 1.The results of microplate semi-quantitative analysis showed that the best inhibitory concentration of agrC specific binding polypeptide was 800 渭 g / ml ~ (2) 路ml ~ (-1) 路L ~ (-1) 路L ~ (-1).The results of microplate semi-quantitative assay showed that when the concentration of specific binding peptide of agrC was 800 渭 g/ml, it could inhibit the formation of bacterial biofilm significantly at 12 h.Scanning electron microscope (SEM) observation showed that the ATCC35984 of Staphylococcus epidermidis was scattered in the small bacterial mass in both the experimental group and the control group at 6 h, and at 12 h, the bacterial mass in the experimental group was significantly lower than that in the control group.At 18 h, the bacterial masses were connected to each other into flakes and tower structures, mature biofilm structures were observed in both groups at 24 h, and a large number of amorphous extracellular matrices were found in the structures. At 30 h, biofilm structures were observed in both groups.But some of them are decomposed.No biofilm formation was observed in the experimental group and control group after culture of Staphylococcus epidermidis ATCC12228 at 612U 1824 and 30h.Laser confocal microscopy showed that the number of bacteria on the surface of PVC in the experimental group was significantly lower than that in the control group, and the number of dead bacteria was more than that in the control group after the addition of specific binding peptide of agrC to the control group and the addition of unrelated peptides to the control group.However, there was no significant difference in bacterial quantity between the two groups at 6: 18 ~ 24 h, and most of them were live bacteria.The bacterial biofilm thickness in the experimental group was significantly lower than that in the control group at 12 h, but there was no significant difference in the other time points.[conclusion] 1.There is a dose-effect relationship between the bacteriostatic strength of Staphylococcus epidermidis and agrC specific binding polypeptide 2.The inhibitory effect of agrC specific binding polypeptide on the formation of biofilm of Staphylococcus epidermidis during the aggregation stage of biofilm formation is significant.Hetero-binding polypeptides can inhibit the formation of staphylococcus epidermidis biofilm. The agrC specific binding peptide can inhibit the further accumulation of Staphylococcus epidermidis.The agglomeration and inhibition of the formation of biofilm can inhibit the proliferation of bacteria and accelerate the death of bacteria during the aggregation stage of biofilm formation.The thickness of bacterial biofilm formation. 3. The specific binding polypeptide of agrC can lay a foundation for the treatment of biofilm formation related infection of Staphylococcus epidermidis, and it may be more effective as a preventive drug.The inhibitory effect on mature biofilm was not obvious.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R515

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