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Title: Typing of Staphylococcus aureus strains by PCR-amplification of variable-length 16S-23S rDNA spacer regions: characterization of spacer sequences.
Austin Authors: Gürtler, V;Barrie, H D
Affiliation: Department of Microbiology, Heidelberg Repatriation Hospital, Victoria, Australia
Issue Date: 1-May-1995
Publication information: Microbiology (reading, England); 141 ( Pt 5)(): 1255-65
Abstract: To develop a rapid and accurate method of typing large numbers of clinical isolates of Staphylococcus aureus, the spacer region C of the rRNA operon [1391-507 (16S-23S)] was enzymically amplified from 322 strains. When the products were separated by denaturing PAGE, 15 variable-length rrn alleles were demonstrated, ranging in size from 906 to 1223 bp. The variable-length HpaII-digested region C [(region E; 1446-196 (16S-23S)] amplification products were cloned into M13mp18RF to sequence separate variable-length alleles. A total of 17 region E inserts were sequenced, aligned and divided into nine alleles by length (938-1174) and sequence properties. The 16S-23S spacer rDNA varied in length (303-551 bp) and in properties; three alleles contained a tRNAIle gene alone, two alleles contained a tRNAIle and a tRNAAla gene, and four alleles lacked tRNA genes. The sequences of two alleles showed less than 1% variation when isolated from two or three S. aureus strains. The 48 penicillin- and methicillin-sensitive strains were divided into 26 ribotypes; in contrast, the 274 methicillin-resistant S. aureus (MRSA) strains were divided into nine ribotypes (A-I) with 97% typing as either ribotype A or B (rrnL was missing in B). The sequence conservation of the rrn operons argues for the use of the 16S-23S spacer region as a stable and direct indicator of the evolutionary divergence of S. aureus strains.
Gov't Doc #: 7773419
Type: Journal Article
Subjects: Alleles
Base Sequence
DNA Primers
DNA, Ribosomal.chemistry.genetics
Genes, Bacterial
Methicillin Resistance.genetics
Molecular Sequence Data
Penicillin Resistance.genetics
Polymerase Chain Reaction.methods
RNA, Ribosomal, 16S.genetics
RNA, Ribosomal, 23S.genetics
RNA, Transfer, Phe.genetics
Reproducibility of Results
Restriction Mapping
Sequence Homology, Nucleic Acid
Staphylococcus aureus.classification.genetics.isolation & purification
Appears in Collections:Journal articles

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