Molecular Diagnosis of Mycoses

对皮肤真菌几丁质合成酶(CHS)基因的DNA序列进行了分析，并保存在基因库中，建立了相应的数据库。对临床分离的皮肤真菌的CHS基因进行了分析，并进行了快速的分子鉴定，证实了临床分离的CHS基因与数据库中的4个DNA序列具有同源性。因此，建立了快速鉴定皮肤真菌的分子方法，并从真菌感染病变的活检标本以及临床血液和排泄物标本中克隆了CHS基因、核糖体DNA、转录间隔区(ITS)范围DNA。并测序，与数据库中报道的DNA序列进行比较。通过这些分子分析，鉴定了包括皮肤癣菌在内的病原性真菌。同时，对这些临床标本的分离株进行了分子检测，证明它们与直接从这些临床标本中获得的结果是一致的。结果也得到了经典鉴定方法的验证。如上所述，与以往常用的方法相比，我们建立的分子方法具有更快速、更简便、灵敏度和特异度都较好的特点。我们一直在开展这一新的分子方法来鉴定出现的临床分离株，并分别诊断动物病例、动物再次出现的真菌病原体和动物真菌病的多样性。所有获得这项研究的人都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.