Development and validation of microarray-based assay for epidemiological study of MRSA.
We have developed a microarray-based assay for the genotyping of Staphylococcus aureus strains. A DNA microarray consisting of 221 genes with 390 oligonucleotide probes was designed to identify characteristic genes or gene alleles of S. aureus. The 221 genes were chosen on the basis of the following criteria: (i) genes used as control for the microarray system, (ii) virulence genes, (iii) resistance genes and their regulators, and (iv) genes constituting genomic islands, e.g., SCCmec. The microarray system was established by determining the method to prepare targets by random-primer labeling with chromosomal DNA and the conditions for hybridization. We verified the system by using DNAs of seven strains, the genome of which has been fully sequenced. Furthermore, the presence of 32 genes and the types of SCCmec elements and coagulase genes carried by another 27 strains were examined and compared with the results of PCR. As a result, the presence or absence of 182 genes out of the 221 genes was verified. Our data showed the usefulness of the oligonucleotide microarray based assay in identifying important marker sets, such as toxin genes, resistance genes, SCCmec elements, and coagulase genes, for the molecular epidemiology of S. aureus.
Otsuka
Junnosuke
J
Probing Technology Laboratory, RIKEN, Wako, Saitama, Japan.
Kondoh
Yasumitsu
Y
Amemiya
Tomoyuki
T
Kitamura
Akio
A
Ito
Teruyo
T
Baba
Satoshi
S
Cui
Longzhu
L
Hiramatsu
Keiichi
K
Tashiro
Tomoko
T
Tashiro
Hideo
H
D015373:Bacterial Typing Techniques; D003030:Coagulase; D015342:DNA Probes / Q000378:metabolism; D005798:Genes, Bacterial; D016106:Methicillin Resistance / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012698:Serologic Tests; D013203:Staphylococcal Infections / Q000453:epidemiology* / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*
D003030:Coagulase; D015342:DNA Probes