Playford E Geoffrey, Kong Fanrong, Sun Ying, Wang Hui, Halliday Catriona, Sorrell Tania C
Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, New South Wales, Australia.
J Clin Microbiol. 2006 Mar;44(3):876-80. doi: 10.1128/JCM.44.3.876-880.2006.
We report on a reverse line blot (RLB) assay, utilizing fungal species-specific oligonucleotide probes to hybridize with internal transcribed spacer 2 region sequences amplified using a nested panfungal PCR. Reference and clinical strains of 16 Candida species (116 strains), Cryptococcus neoformans (five strains of Cryptococcus neoformans var. neoformans, five strains of Cryptococcus neoformans var. grubii, and six strains of Cryptococcus gatti), and five Aspergillus species (68 strains) were all correctly identified by the RLB assay. Additional fungal species (16 species and 26 strains) not represented on the assay did not exhibit cross-hybridization with the oligonucleotide probes. In simulated clinical specimens, the sensitivity of the assay for Candida spp. and Aspergillus spp. was 10(0.5) cells/ml and 10(2) conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens.
我们报告了一种反向线印迹(RLB)检测方法,该方法利用真菌物种特异性寡核苷酸探针与通过巢式泛真菌PCR扩增的内转录间隔区2区域序列进行杂交。16种念珠菌(116株)、新型隐球菌(新型隐球菌变种新型隐球菌5株、新型隐球菌变种格鲁比隐球菌5株和加氏隐球菌6株)以及5种曲霉菌(68株)的参考菌株和临床菌株均通过RLB检测得到正确鉴定。检测方法中未涵盖的其他真菌物种(16种和26株)未与寡核苷酸探针发生交叉杂交。在模拟临床标本中,该检测方法对念珠菌属和曲霉菌属的敏感性分别为10(0.5) 个细胞/毫升和10(2) 个分生孢子/毫升。该检测方法能够灵敏且特异同时检测和鉴定多种真菌病原体。