Intra-Genomic Internal Transcribed Spacer Region Sequence Heterogeneity and Molecular Diagnosis in Clinical Microbiology

International Journal of Molecular Sciences

Yingchun Xu¹  Meng Xiao¹  Jingwei Cheng¹  Ying Zhao¹  Chi-Ching Tsang²  Susanna K. P. Lau²  Patrick C. Y. Woo² 

[1] Department of Clinical Laboratory, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences, Beijing 100730, China;Department of Microbiology, The University of Hong Kong, Hong Kong;

关键词: internal transcribed spacer region;    sequencing;    heterogeneity;    yeast;    molecular identification;   

DOI  :  10.3390/ijms161025067

来源: DOAJ

【 摘 要 】

Internal transcribed spacer region (ITS) sequencing is the most extensively used technology for accurate molecular identification of fungal pathogens in clinical microbiology laboratories. Intra-genomic ITS sequence heterogeneity, which makes fungal identification based on direct sequencing of PCR products difficult, has rarely been reported in pathogenic fungi. During the process of performing ITS sequencing on 71 yeast strains isolatedfrom various clinical specimens, direct sequencing of the PCR products showedambiguous sequences in six of them. After cloning the PCR products into plasmids for sequencing, interpretable sequencing electropherograms could be obtained. For each ofthe six isolates, 10–49 clones were selected for sequencing and two to seven intra-genomic ITS copies were detected. The identities of these six isolates were confirmed to beCandida glabrata (n = 2), Pichia (Candida) norvegensis (n = 2), Candida tropicalis (n = 1) and Saccharomyces cerevisiae (n = 1). Multiple sequence alignment revealed that one to four intra-genomic ITS polymorphic sites were present in the six isolates, and all these polymorphic sites were located in the ITS1 and/or ITS2 regions. We report and describe the first evidence of intra-genomic ITS sequence heterogeneity in four different pathogenic yeasts, which occurred exclusively in the ITS1 and ITS2 spacer regions for the six isolates in this study.

【 授权许可】

Unknown