巢式PCR扩增诊断播散性毛孢子菌病的动物实验研究  

A nested PCR assay to diagnose disseminated trichosporonosis in murine model

作　　者：杨蓉娅[1] 赵逊[1] 郝飞[1] 

机构地区：[1]第三军医大学西南医院皮肤科,重庆400038

出　　处：《第三军医大学学报》2006年第14期1506-1508,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目(30570099)~~

摘　　要：目的探讨应用PCR技术诊断播散性毛孢子菌病的可行性。方法建立动物模型后,于接种3、7、14 d分别采集小鼠眶静脉血和肝脏组织标本,进行真菌培养和巢式PCR扩增。结果小鼠静脉接种3、7、14 d后血清培养的阳性率分别为0、20%、0,而巢式PCR的阳性率分别为80%、90%、88.9%;在上述时相点,肝脏组织真菌培养阳性率分别为50%、40%、22.2%,而巢式PCR的阳性率分别为90%、90%、88.9%。血清和肝脏组织真菌培养阳性率与巢式PCR阳性率差异均具有统计学意义(P<0.05,P<0.01)。结论巢式PCR扩增T.asahii特异性DNA片段,可作为播散性毛孢子菌病的一种特异、灵敏、快捷的早期诊断手段。Objective To explore the feasibility of the nested PCR technique to detect the Trichosporon asahii DNA in the blood and liver samples from the murine model of disseminated trichosporonosis. Methods On day 3, 7, 14 after intravenous inoculation of Trichosporon asahii suspension, blood and liver samples of 30 BALB/c mice were collected. Another ten normal mice serve as control. DNA was amplified by nested PCR, with a pair of primer. Fungal culture was done as control. Results On day 3, 7, 14 after inoculation, the positive rate of the serum fungal cultivation were 0, 20%, 0 ; the positive rate of the serum after nested PCR assay was 80%, 90%, 88.9% ; the positive rate of liver fungal cultivation were 50%, 40%, 22.2% ; the positive rate of liver after nested PCR assay was 90%, 90%, 88.9%. There were statistical differences between the positive rate by nested PCR assay and fungal cultivation to check the serum and liver samples on day 3, 7, 14 after inoculation. The survival mice to the 14th day were too less to make statistical analysis. Conclusion The results suggest that T. asahii DNA assay by nested PCR is more specific, more sensitive, and faster than fungal cultivation. The nested PCR assay could be a potential assay for the diagnosis of disseminated trichosporonosis.

关 键 词：播散性毛孢子菌病 诊断 

分 类 号：R446.5[医药卫生—诊断学] R519.8[医药卫生—临床医学]