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Authors:Chen,Y.C.;Eisner,J.D.;Kattar,M.M.;Rassoulian-Barrett,S.L.;Lafe,K.;Bui,U.;Limaye,A.P.;Cookson,B.T. 
Year:2001 
Journal:
 
Volume:39 
Keywords:GENBANK/AF335925 GENBANK/AF335926 GENBANK/AF335927 GENBANK/AF335928 GENBANK/AF335929 GENBANK/AF335930 GENBANK/AF335931 GENBANK/AF335932 GENBANK/AF335933 GENBANK/AF335934 GENBANK/AF335935 GENBANK/AF335936 GENBANK/AF335937 GENBANK/AF335938 GENBANK/AF335939 GENBANK/AF335940 GENBANK/AF335941 GENBANK/AF335942 GENBANK/AF335943 GENBANK/AF335944 GENBANK/AF335945 GENBANK/AF335946 GENBANK/AF335947 GENBANK/AF335948 GENBANK/AF335949 GENBANK/AF335950 GENBANK/AF335951 GENBANK/AF335952 GENBANK/AF335953 GENBANK/AF335954 GENBANK/AF335955 GENBANK/AF335956 GENBANK/AF335957 GENBANK/AF335958 GENBANK/AF335959 GENBANK/AF335960 GENBANK/AF335961 GENBANK/AF335962 GENBANK/AF335971 GENBANK/AF335972 GENBANK/AF335973 GENBANK/AF335974 GENBANK/AF335975 GENBANK/AF335976 GENBANK/AF335977 GENBANK/AF335978 GENBANK/AF335979 GENBANK/AF335980 GENBANK/AF335981 GENBANK/AF335982 GENBANK/AF335983 GENBANK/AF335984 GENBANK/AF335985 GENBANK/AF335986 GENBANK/AF335987 GENBANK/AF335988 GENBANK/AF336831 GENBANK/AF336832 GENBANK/AF336833 GENBANK/AF336834 GENBANK/AF336835 GENBANK/AF336836 GENBANK/AF336837 GENBANK/AF336838 GENBANK/AF336839 GENBANK/AF336840 GENBANK/AF336841 GENBANK/AF336842 GENBANK/AF336843 GENBANK/AF336844 GENBANK/AF336845 GENBANK/AF336846 DNA, Fungal/analysis DNA, Ribosomal Spacer/*analysis Databases, Factual Electrophoresis, Capillary Human Molecular Sequence Data Mycoses/*microbiology Phylogeny Polymerase Chain Reaction Polymorphism (Genetics) RNA, Ribosomal/genetics Sequence Analysis, DNA Yeasts/*classification/*genetics 
Abstract:Species-specific polymorphisms in the noncoding internal transcribed spacer 2 (ITS2) region of the rRNA operon provide accurate identification of clinically significant yeasts. In this study, we tested the hypothesis that ITS1 noncoding regions contain diagnostically useful alleles. The length of ITS1 region PCR products amplified from 40 species (106 clinical strains, 5 reference strains, and 30 type strains) was rapidly determined with single-base precision by automated capillary electrophoresis. Polymorphisms in the PCR product length permitted 19 species to be distinguished by ITS1 alone, compared with 16 species distinguished by using only ITS2. However, combination of both ITS alleles permitted identification of 30 species (98% of clinical isolates). The remaining 10 species with PCR products of similar sizes contained unique ITS alleles distinguishable by restriction enzyme analysis. DNA sequence analysis of amplified ITS1 region DNA from 79 isolates revealed species-specific ITS1 alleles for each of the 40 pathogenic species examined. This provided identification of unusual clinical isolates, and 53 diagnostic ITS1 sequences were deposited in GenBank. Phylogenetic analyses based on ITS sequences showed a similar overall topology to 26S rRNA gene-based trees. However, different species with identical 26S sequences contained distinct ITS alleles that provided species identification with strong statistical support. Together, these data indicate that the analysis of ITS polymorphisms can reliably identify 40 species of clinically significant yeasts and that the capacity for identifying potentially new pathogenic species by using this database holds significant promise. 
Summary:Chen,Y.C.;Eisner,J.D.;Kattar,M.M.;Rassoulian-Barrett,S.L.;Lafe,K.;Bui,U.;Limaye,A.P.;Cookson,B.T. 2001. Journal of Clinical Microbiology. 39 
Page:4042-51 
Num_:11 
ISSN:0095-1137 
Revue:1627 
Url:11682528 
Document Reference #:9614