Molecular Diagnosis of Mycoses

研究了皮肤植物几丁质合成酶（CHS）基因的DNA序列，并将其存入基因库，建立了数据库。对临床分离的皮肤癣菌CHS基因进行了分析，并进行了快速分子鉴定，确认了临床分离株的DNA序列与数据库中的DNA序列具有同源性。为此，建立了快速分子鉴别皮肤真菌的方法。此外，从真菌感染病变的活检标本以及临床血液和排泄物样本中，克隆了CHS基因、核糖体DNA、转录间隙（ITS）范围DNA。和测序，以比较数据库中报告的DNA序列。通过这些分子分析，鉴定了包括皮癣菌在内的病原真菌。同时，从这些临床标本中分离的菌株也进行了分子检测，证明它们与直接从这些临床标本中获得的结果一致。经典鉴定方法也证实了这一结果。如上所述，我们开发的分子方法在诊断真菌感染方面比以往一般使用的方法更快速、更容易、灵敏度和特异性都好。我们一直在开展这种新的分子方法来鉴定临床分离株，分别诊断动物病例、动物中再出现的真菌病原体和动物真菌病的多样性。所有获得的研究成果均已在国际期刊上发表。

The DNA sequences of chitin synthase (CHS) genes in dermatophytes were elucidated, and deposited on the Gene Bank, developing the data base. CHS genes in clinical isolates of dermatophytes were also analyzed and a rapid molecular identification was performed confirming the homology between the DNA sequences of clinical isolates an4 those on the data base. Therefore, the rapid molecular method was established for the identification of dermatophytes.Furthermore, from biopsy specimens from the lesions of fungal infection, as well as clinical samples of blood, and excretions, CHS genes, ribosomal DNA, intertranscribed space (ITS) range DNA were cloned. and sequenced ,to compare the DNA sequences reported on data base. By these molecular analysis, etiologic fungi including dermatophytes were identified.Simultaneously, the isolates from these clinical samples were also examined molecularly proving that they were consisting to the results obtained directly from these clinical specimens. The results were also confirmed by classical identification methods.As mentioned above, the molecular method we developed was evaluated to be more rapid and easier with good quality in sensitivity and in specificity for the diagnosis of fungal infection rather than the previous method used in general. We have been carrying out this new molecular method to identify the clinical isolates emerging and to diagnose the animal cases, re-emerging fungal pathogens in animals, respectively and the diversity of animal mycoses. All obtained this research were already published on the international journals.