Document Detail


Practical identification of eight medically important Trichosporon species by reverse line blot hybridization (RLB) assay and rolling circle amplification (RCA).
MedLine Citation:
PMID:  23186014     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We developed a reverse line blot (RLB) hybridization-, and rolling circle amplification (RCA)-based assays for the identification of Trichoporon species and evaluated them with 48 isolates that had been previously recognized as belonging to eight species (Trichosporon asahii, T. cutaneum, T. dermatis, T. domesticum, T. inkin, T. japonicum, T. jirovecii, and T. laibachii). Results were compared to those obtained with DNA sequencing of three rRNA gene loci, i.e., the internal transcribed spacer (ITS) region, D1/D2 domain of the 28S rRNA gene and intergenic spacer 1 (IGS1) region. Using species-specific, or group-specific probes targeted at the ITS region and the D1/D2 domain, the RLB assay permitted accurate species identification of all 48 isolates with 100% specificity. Species-specific RLB probes correctly assigned 45/48 (94%) of the isolates (six species) with the exception of T. dermatis and T. japonicum isolates which were not targeted by the assay. Identification of T. dermatis relied on a positive hybridization result with the group-specific probe hybridizing with T. dermatis and T. jirovecii and the absence of a signal with the T. jirovecii-specific probe. T. japonicum strains were first assigned to the T. asahii-T. japonicum group by hybridization with the two species group-specific probe and then as T. japonicum by the absence of signal with a T. asahii-specific probe. Twelve species-specific RCA probes targeting the eight species studied detected templates of all 48 Trichosporon isolates and an artificial template of T. asteroides, all with good specificity. Both RLB and RCA are potential alternatives to DNA sequencing for the identification of Trichosporon species. The RLB approach is suited for the batched simultaneous analysis of large numbers of isolates, while RCA is more appropriate for the immediate study of single isolates. Comparative costs are US$7 and US$2 per assay for the RLB and RCA methods, respectively.
Authors:
Meng Xiao; Li-Na Guo; Fanrong Kong; He Wang; Tania C Sorrell; Ruo-Yu Li; Wei Jiang; Sharon C-A Chen; Ying-Chun Xu
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-11-28
Journal Detail:
Title:  Medical mycology     Volume:  51     ISSN:  1460-2709     ISO Abbreviation:  Med. Mycol.     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-03-18     Completed Date:  2013-12-11     Revised Date:  2013-12-13    
Medline Journal Info:
Nlm Unique ID:  9815835     Medline TA:  Med Mycol     Country:  England    
Other Details:
Languages:  eng     Pagination:  300-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Humans
Microbiological Techniques / methods*
Mycology / methods*
Nucleic Acid Amplification Techniques / methods*
Nucleic Acid Hybridization / methods*
Oligonucleotide Probes / genetics
Sensitivity and Specificity
Trichosporon / classification*,  genetics,  isolation & purification*
Chemical
Reg. No./Substance:
0/Oligonucleotide Probes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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