应用巢式聚合酶链反应(Nested PCR)—限制片段长度多态性(Restriction fragment length polymorphism,RFLP)方法对微小隐孢子虫(Cryptosporidiumparvum)、安氏隐孢子虫(C.andersoni)、贝氏隐孢子虫(C.baileyi)和猪隐孢子虫(C.suis)的鉴...应用巢式聚合酶链反应(Nested PCR)—限制片段长度多态性(Restriction fragment length polymorphism,RFLP)方法对微小隐孢子虫(Cryptosporidiumparvum)、安氏隐孢子虫(C.andersoni)、贝氏隐孢子虫(C.baileyi)和猪隐孢子虫(C.suis)的鉴别进行了研究。结果显示:牛源C.parvum、羊源C.parvum、C.suis、C.andersoni和C.baileyi扩增产物片段大小分别为212,213,213,213,210 bp;扩增产物先后经TaqⅠ、BstUⅠ与MseⅠ酶切,根据酶切后形成的不同酶切图谱可以鉴别C.parvum、C.suis、C.andersoni和C.baileyi。该研究为我国隐孢子虫的分类和隐孢子虫病的分子流行病学调查提供了新的方法。展开更多
Summary: To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFL...Summary: To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting TopoisomeraseⅡgene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme HincⅡ. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of HincⅡ, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase Ⅱ gene is rapid and efficient.展开更多
文摘Summary: To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting TopoisomeraseⅡgene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme HincⅡ. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of HincⅡ, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase Ⅱ gene is rapid and efficient.
