【摘要】： 目的:大鼠、小鼠、豚鼠、家兔等作為實驗動物,已廣泛應用于科研、教學和生產實踐中。三種致病性皮膚真菌:犬小孢子菌( Microsporum canis )、石膏樣小孢子菌(Microsprum gypseum)、須毛癬菌( Trichonohyton mentagrophytes),是所有實驗動物應排除的病原菌,不論是從動物皮膚病灶處或從非病灶處分離到這三種菌,均視為異常。因為非病灶處的真菌很可能隨后引起實驗動物的皮膚病,并有可能造成飼養(yǎng)和實驗人員的感染。 目前該病的實驗室診斷主要依賴于形態(tài)學觀察(直接鏡檢及真菌培養(yǎng)),但所需時間長,且不易鑒定到種的水平,尤其需要較長的工作經驗,因而不能滿足實驗動物快速、準確檢測的需要。近年來,分子生物學方法開始應用于真菌學研究方面,如RFLP、分子雜交、DNA序列分析等,特別是任意引物聚合酶鏈式反應(AP-PCR),有力地促進了皮膚病原真菌檢測的發(fā)展。本研究將皮膚病原真菌通用引物PCR與AP-PCR技術相結合,應用于實驗動物皮膚病原真菌的檢測,使真菌的表型鑒定轉向基因型鑒定,以達到快速準確檢測實驗動物皮膚病原真菌的目的。 方法: 1標準菌株的DNA提取及PCR擴增:取標準菌株犬小孢子菌、石膏樣小孢子菌、須毛癬菌傳種至沙氏斜面固體培養(yǎng)基,27℃培養(yǎng)7天。無菌條件下各取少量以上三種培
[Abstract]:Objective: as experimental animals, rats, mice, guinea pigs and rabbits have been widely used in scientific research, teaching and production. Three kinds of pathogenic dermatophytes: (Microsporum canis), (Microsprum gypseum), (Trichonohyton mentagrophytes), are the pathogens that should be excluded from all experimental animals, whether they are isolated from the lesions of animal skin or from non-foci. Are considered abnormal. Because nonfocal fungi are likely to subsequently cause dermatosis in laboratory animals, and may cause infection in breeding and laboratory workers. At present, the laboratory diagnosis of the disease mainly depends on morphological observation (direct microscopic examination and fungal culture), but it takes a long time, and it is difficult to identify the level of the species, especially requires a long working experience, so it can not meet the needs of laboratory animals quickly. The need for accurate detection. In recent years, molecular biological methods have been applied to mycological research, such as RFLP, hybridization, DNA sequence analysis, especially arbitrary primer polymerase chain reaction (AP-PCR), which have promoted the development of dermatogenic fungi detection. In this study, the general primer PCR and AP-PCR technique were used to detect dermatogenic fungi in laboratory animals, and the phenotypic identification of fungi was turned to genotyping. In order to achieve the purpose of rapid and accurate detection of dermatogenic fungi in laboratory animals. Methods: (1) DNA extraction and PCR amplification of standard strains: microspora canis, microsporum gypsum, Trichophyton tuber were transferred to solid culture medium and cultured at 27 鈩，

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