Latouche G N, Daniel H M, Lee O C, Mitchell T G, Sorrell T C, Meyer W
Centre for Infectious Diseases and Microbiology, The University of Sydney, Westmead Hospital, Australia.
J Clin Microbiol. 1997 Dec;35(12):3171-80. doi: 10.1128/jcm.35.12.3171-3180.1997.
A total of 49 type and neotype isolates and 32 clinical isolates of the anamorph genus Candida and related teleomorph genera were obtained from different culture collections and clinical laboratories. Isolates were subjected to two phenotypic methods of identification, Vitek yeast biochemical card (YBC) and API ID 32C, both based on carbohydrate assimilation, and one genotypic method, PCR fingerprinting, based on the detection of DNA polymorphisms between minisatellite-specific sequences with the primer M13 (5' GAGGGTGGCGGTTCT 3'). The correct identification of a strain at the Centraalbureau voor Schimmelcultures was used as the gold standard for the identification of an isolate. When the study was restricted to species included in the respective biochemical databases, the Vitek YBC and API ID 32C systems performed adequately with positive identification rates of 87.3 and 76.8%, respectively. When uncommon species were added to the study, several of which are not included in the databases, the identification efficiencies were 76.5 and 77.5%, respectively. By comparison, all isolates were correctly identified by PCR fingerprinting, with 63 reference species profiles in the databank. Sufficient polymorphisms among the total set of banding patterns were observed, with adequate similarity in the major patterns obtained from a given species, to allow each isolate to be assigned unambiguously to a particular species. In addition, variations in minor bands allowed for differentiation to the strain level. PCR fingerprinting was found to be rapid, reproducible, and more cost-effective than either biochemical approach. Our results provide reference laboratories with an improved identification method for yeasts based on genotypic rather than phenotypic markers.
从不同的菌种保藏中心和临床实验室共获得了49株形态型和新模式菌株以及32株无性型念珠菌属和相关有性型属的临床分离株。分离株采用两种基于碳水化合物同化作用的表型鉴定方法,即Vitek酵母生化鉴定卡(YBC)和API ID 32C,以及一种基于用引物M13(5' GAGGGTGGCGGTTCT 3')检测小卫星特异性序列之间DNA多态性的基因型方法,即PCR指纹图谱法。以荷兰真菌保藏中心对菌株的正确鉴定作为分离株鉴定的金标准。当研究仅限于各自生化数据库中包含的物种时,Vitek YBC和API ID 32C系统表现良好,阳性鉴定率分别为87.3%和76.8%。当将一些数据库中未包含的罕见物种纳入研究时,鉴定效率分别为76.5%和77.5%。相比之下,通过PCR指纹图谱法所有分离株均被正确鉴定,数据库中有63种参考物种图谱。在整个条带模式集中观察到了足够的多态性,从给定物种获得的主要模式具有足够的相似性,以便能够将每个分离株明确地归为特定物种。此外,次要条带的差异允许区分到菌株水平。结果发现,PCR指纹图谱法快速、可重复,且比任何一种生化方法都更具成本效益。我们的结果为参考实验室提供了一种基于基因型而非表型标记的改进的酵母鉴定方法。