Elie C M, Lott T J, Reiss E, Morrison C J
Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
J Clin Microbiol. 1998 Nov;36(11):3260-5. doi: 10.1128/JCM.36.11.3260-3265.1998.
Rapid identification of Candida species has become more important because of an increase in infections caused by species other than Candida albicans, including species innately resistant to azole antifungal drugs. We previously developed a PCR assay with an enzyme immunoassay (EIA) format to detect amplicons from the five most common Candida species by using universal fungal primers and species-specific probes directed to the ITS2 region of the gene for rRNA. We designed probes to detect seven additional Candida species (C. guilliermondii, C. kefyr, C. lambica, C. lusitaniae, C. pelliculosa, C. rugosa, and C. zeylanoides) included in the API 20C sugar assimilation panel, five probes for species not identified by API 20C (C. haemulonii, C. norvegica, C. norvegensis, C. utilis, and C. viswanathii), and a probe for the newly described species C. dubliniensis, creating a panel of 18 Candida species probes. The PCR-EIA correctly identified multiple strains of each species tested, including five identified as C. albicans by the currently available API 20C database but determined to be C. dubliniensis by genotypic and nonroutine phenotypic characteristics. Species identification time was reduced from a mean of 3.5 days by conventional identification methods to 7 h by the PCR-EIA. This method is simple, rapid, and feasible for identifying Candida species in clinical laboratories that utilize molecular identification techniques and provides a novel method to differentiate the new species, C. dubliniensis, from C. albicans.
由于非白色念珠菌引起的感染有所增加,包括对唑类抗真菌药物天然耐药的菌种,快速鉴定念珠菌属变得更为重要。我们之前开发了一种采用酶免疫测定(EIA)形式的PCR检测方法,通过使用通用真菌引物和针对rRNA基因ITS2区域的种特异性探针,来检测五种最常见念珠菌属菌种的扩增子。我们设计了探针来检测API 20C糖同化鉴定板中包含的另外七种念珠菌属菌种(季也蒙念珠菌、克柔念珠菌、羔羊念珠菌、葡萄牙念珠菌、膜状念珠菌、皱落念珠菌和锡兰念珠菌),五种用于检测API 20C未能鉴定的菌种(哈氏念珠菌、挪威念珠菌、新挪威念珠菌、解脂念珠菌和维斯瓦那斯念珠菌)的探针,以及一种用于新描述的菌种都柏林念珠菌的探针,从而创建了一个包含18种念珠菌属菌种的探针组。PCR-EIA正确鉴定了所检测的每种菌种的多个菌株,包括目前可用的API 20C数据库鉴定为白色念珠菌,但根据基因型和非常规表型特征确定为都柏林念珠菌的五个菌株。菌种鉴定时间从传统鉴定方法的平均3.5天缩短至PCR-EIA的7小时。该方法对于利用分子鉴定技术进行临床实验室念珠菌属菌种鉴定而言简单、快速且可行,并提供了一种区分新菌种都柏林念珠菌与白色念珠菌的新方法。