实验动物皮肤病原真菌分子生物学检测方法的建立及应用
发布时间：2018-10-21 13:29
【摘要】： 目的:大鼠、小鼠、豚鼠、家兔等作为实验动物,已广泛应用于科研、教学和生产实践中。三种致病性皮肤真菌:犬小孢子菌( Microsporum canis )、石膏样小孢子菌(Microsprum gypseum)、须毛癣菌( Trichonohyton mentagrophytes),是所有实验动物应排除的病原菌,不论是从动物皮肤病灶处或从非病灶处分离到这三种菌,均视为异常。因为非病灶处的真菌很可能随后引起实验动物的皮肤病,并有可能造成饲养和实验人员的感染。 目前该病的实验室诊断主要依赖于形态学观察(直接镜检及真菌培养),但所需时间长,且不易鉴定到种的水平,尤其需要较长的工作经验,因而不能满足实验动物快速、准确检测的需要。近年来,分子生物学方法开始应用于真菌学研究方面,如RFLP、分子杂交、DNA序列分析等,特别是任意引物聚合酶链式反应(AP-PCR),有力地促进了皮肤病原真菌检测的发展。本研究将皮肤病原真菌通用引物PCR与AP-PCR技术相结合,应用于实验动物皮肤病原真菌的检测,使真菌的表型鉴定转向基因型鉴定,以达到快速准确检测实验动物皮肤病原真菌的目的。 方法: 1标准菌株的DNA提取及PCR扩增:取标准菌株犬小孢子菌、石膏样小孢子菌、须毛癣菌传种至沙氏斜面固体培养基,27℃培养7天。无菌条件下各取少量以上三种培
[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，
本文编号：2285216
[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，
本文编号：2285216
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2285216.html
最近更新
教材专著
