葡萄孢弗隆那霉致病性的小鼠模型和基因型研究  

Experimental study on genotypes and pathogenicity of Veronaeae botryosa in mice

作　　者：王红[1] 吕桂霞[3] 张燕[2] 魏欣净[1] 胡素泉[3] 沈永年[3] 刘维达[3] 

机构地区：[1]山东大学附属省立医院皮肤科,济南250021 [2]滨州医学院病理学教研室 [3]中国医学科学院北京协和医学院皮肤病研究所

出　　处：《中华皮肤科杂志》2009年第8期533-536,共4页Chinese Journal of Dermatology

基　　金：公益性卫生行业科研专项经费项目（200802032） 志谢 本实验中的河南株获赠于北京大学第一医院医学真菌中心

摘　　要：目的探讨两株具有不同最高生长温度的葡萄孢弗隆那霉临床分离株的致病力、基因型的差异。方法将在37℃生长良好的江苏株和在36℃不生长的河南株葡萄孢弗隆那霉造成实验性感染，观察动物的发病、死亡以及各脏器受累情况。提取两临床分离株的DNA进行随机扩增多态DNA（RAPD）分析以及核糖体内转录间隔区（ITS区）扩增，并对产物测序。结果江苏株可造成免疫正常及免疫抑制小鼠的暗色丝孢霉病，前者死亡率为30％，后者死亡率65％，两组间差异有统计学意义。免疫抑制组小鼠的肺脏感染率为100％，明显高于其他脏器，差异具有统计学意义；肝脏直接镜检阳性率为64．7％，组织培养阳性率为70．5％；免疫正常组各脏器感染阳性率差异无统计学意义，肺脏阳性率为57．8％，肝脏为42．1％。免疫抑制组小鼠的肺脏感染率明显高于免疫正常组。河南株在免疫正常组和免疫抑制组均未见导致明确的感染。RAPD和ITS区测序结果显示，在ITS区的236位碱基处有一个碱基的差异（A—G）。结论两株菌具有不同的基因型；葡萄孢弗隆那霉江苏株可以导致免疫抑制和免疫正常小鼠的系统性暗色丝孢霉病，前者的致死率显著高于后者，致病力与其免疫状态有关；免疫抑制状态下，肺脏是其最常受累的器官。Objective To study the difference in pathogenicity and genotype between two isolates of Veronaeae botryosa with different temperature tolerance. Methods Two strains of Veronaeae botryose were isolated from two patients with phaeohyphomycosis in Jiangsu and Henan province respectively. Of them, the Jiangsu strain could grow well at 37℃, but Henan strain could not grow at 36 ℃. Eighty mice were equally classified into immunocompetent and immuno-suppressed （induced by cyclophosphamide） groups to be inoculated with the two strains of Veronaeae botryosa respectively. Ten mice remained uninoculated and served as the control. The general condition, growth and organic involvement of mice were observed for 4 weeks followed by the killing of surviving mice. Homogenated tissue samples were obtained from liver, spleen, lung, kidney and brain; then, tissue culture, direct microscopy and pathological examination were performed. Genomic DNA was extracted from tissue samples and subjected to random amplified polymorphic DNA （RAPD） analysis. PCR was performed to amplify the intemal transcribed spacer （ITS） ofrDNA followed by sequencing. Results Systemic phaeohyphomycosis was induced in both immunocompetent and immuno-suppressed mice by the Jiangsu strain of Veronaeae botryose; the mortality was 30% in immuno- competent mice and 65% in immuno-suppressed mice with statistical significance between the two groups. In immuno-suppressed mice inoculated with the Jiangsu strain, the infection rate was 100% in the lung, signifi- cantly higher than in other organs; on direct microscopy the infection rate reached 64.7% in the liver, and 70.5% on tissue culture. There was no significant difference in the infection rate among these organs in immunocompetent mice inoculated with the Jiangsu strain, with the infection rate being 57.8% in the lung and 42.1% in the liver. Increased infection rate was observed in the lung of immuno-suppressed mice compared with immunocompetent mice （P 〈 0.05 ）. No definite infection was seen

关 键 词：葡萄孢弗隆那霉 毒力 模型 动物 

分 类 号：R-332[医药卫生]