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地塞米松对红色毛癣菌生物学性状的影响

发布时间:2019-06-11 09:25
【摘要】:目的:1.观察地塞米松对红色毛癣菌生长、菌落产色及抗真菌药物敏感性的影响,为临床及时准确的诊断和合理的用药提供理论依据。2.探讨红色毛癣菌在地塞米松干预前后基因型的差异,从基因水平探讨地塞米松影响下红色毛癣菌表型特征发生变化的可能分子生物学机制。 方法:1.对临床上经分离鉴定的红色毛癣菌首先采用大培养测定含不同浓度地塞米松沙氏培养基上菌落生长直径,并绘制生长曲线,然后采用分光光度计测定含不同浓度地塞米松沙氏液体培养基的吸光度,最后参照美国国家实验室标准委员会(NCCLS M-38A)方案,采用微量液基法检测不同浓度地塞米松干预下特比萘芬对红色毛癣菌的MIC变化。2.取在含不同浓度地塞米松培养基上生长的红色毛癣菌,采用真菌通用引物ITS1、ITS4对其ITS区进行PCR扩增,从分子水平对其进行鉴定,经鉴定的红色毛癣菌进一步对其NTS区进行PCR扩增,观察不同浓度地塞米松干预前后基因条带的变化。 结果:1.从绘制的生长曲线看,当地塞米松的浓度为0.05%、0.1%和0.2%时,与对照组比较无明显的变化,当地塞米松的浓度为0.4%、0.8%时前五天对红色毛癣菌生长有明显的抑制作用,其后促进红色毛癣菌的生长,当地塞米松的浓度为1.0%时对红色毛癣菌生长有明显的抑制作用。2.不含地塞米松组红色毛癣菌的吸光度值(OD)几何均数为0.117±0.032,特比萘芬MIC为0.063μg/ml,当地塞米松的浓度为0.05%、0.1%和0.2%时,,红色毛癣菌的OD几何均数分别为0.118±0.032、0.118±0.032和0.132±0.030,MIC分别为0.081μg/ml、0.092μg/ml和0.109μg/ml与空白组比较无明显差异(P>0.05);当其浓度为0.4%、0.8%时,红色毛癣菌的OD几何均数分别为0.165±0.081、0.175±0.085,特比萘芬MIC分别为0.196μg/ml、0.319μg/ml与空白组比较有明显的差异(P<0.05)。3.引物ITS1、ITS4在不同浓度地塞米松沙氏培养基上生长的红色毛癣菌均扩增出一约690bp特异条带,5株红色毛癣菌用引物TrNTSF-2和
[Abstract]:Objective: 1. To observe the effect of dexamethasone on the growth, colony color production and antifungal drug sensitivity of Trichophyton rubrum. To provide theoretical basis for timely and accurate diagnosis and reasonable drug use in clinic. 2. To explore the genotypic difference of Trichophyton rubrum before and after dexamethasone intervention. To explore the possible molecular biological mechanism of phenotypic changes of Trichophyton rubrum under the influence of dexamethasone at gene level. Methods: 1. The colony growth diameter of Trichophyton rubrum isolated and identified in clinic was determined by large culture, and the growth curve was drawn. Then the absorbance of the liquid medium containing different concentrations of dexamethasone was determined by spectrophotometer. Finally, the National Laboratory Standards Committee (NCCLS M 鈮

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