Combination of Multiplex PCRs for Staphylococcal Cassette Chromosome mec Type Assignment: Rapid Identification System for mec, ccr, and Major Differences in Junkyard Regions
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| Published in | Antimicrobial Agents and Chemotherapy Vol. 51; no. 1; pp. 264 - 274 |
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| Language | English |
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American Society for Microbiology
01.01.2007
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| AbstractList | Staphylococcal cassette chromosome
mec
(SCC
mec
) typing, in combination with genotyping of the
Staphylococcus aureus
chromosome, has become essential for defining methicillin-resistant
S. aureus
(MRSA) clones in epidemiological studies. We have developed a convenient system for SCC
mec
type assignment. The system consists of six multiplex PCRs (M-PCRs) for identifying the
ccr
gene complex (
ccr
), the
mec
gene complex (
mec
), and specific structures in the junkyard (J) regions: M-PCR with primer set 1 (M-PCR 1) identified five types of
ccr
genes; M-PCR 2 identified class A to class C
mec
; M-PCRs 3 and 4 identified specific open reading frames in the J1 regions of type I and IV and of type II, III, and V SCC
mec
elements, respectively; M-PCR 5 identified the transposons Tn
554
and ΨTn
554
integrated into the J2 regions of type II and III SCC
mec
elements; and M-PCR 6 identified plasmids pT181 and pUB110 integrated into J3 regions. The system was validated with 99 MRSA strains carrying SCC
mec
elements of different types. The SCC
mec
types of 93 out of the 99 MRSA strains could be assigned. The SCC
mec
type assignments were identical to those made with a PCR system that uses numerous primer pairs to identify genes or gene alleles. Our system of six M-PCRs is thus a convenient and reliable method for typing SCC
mec
elements. Staphylococcal cassette chromosome mec (SCCmec) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for defining methicillin-resistant S. aureus (MRSA) clones in epidemiological studies. We have developed a convenient system for SCCmec type assignment. The system consists of six multiplex PCRs (M-PCRs) for identifying the ccr gene complex (ccr), the mec gene complex (mec), and specific structures in the junkyard (J) regions: M-PCR with primer set 1 (M-PCR 1) identified five types of ccr genes; M-PCR 2 identified class A to class C mec; M-PCRs 3 and 4 identified specific open reading frames in the J1 regions of type I and IV and of type II, III, and V SCCmec elements, respectively; M-PCR 5 identified the transposons Tn554 and PsiTn554 integrated into the J2 regions of type II and III SCCmec elements; and M-PCR 6 identified plasmids pT181 and pUB110 integrated into J3 regions. The system was validated with 99 MRSA strains carrying SCCmec elements of different types. The SCCmec types of 93 out of the 99 MRSA strains could be assigned. The SCCmec type assignments were identical to those made with a PCR system that uses numerous primer pairs to identify genes or gene alleles. Our system of six M-PCRs is thus a convenient and reliable method for typing SCCmec elements. Staphylococcal cassette chromosome mec (SCCmec) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for defining methicillin-resistant S. aureus (MRSA) clones in epidemiological studies. We have developed a convenient system for SCCmec type assignment. The system consists of six multiplex PCRs (M-PCRs) for identifying the ccr gene complex (ccr), the mec gene complex (mec), and specific structures in the junkyard (J) regions: M-PCR with primer set 1 (M-PCR 1) identified five types of ccr genes; M-PCR 2 identified class A to class C mec; M-PCRs 3 and 4 identified specific open reading frames in the J1 regions of type I and IV and of type II, III, and V SCCmec elements, respectively; M-PCR 5 identified the transposons Tn554 and psi Tn554 integrated into the J2 regions of type II and III SCCmec elements; and M-PCR 6 identified plasmids pT181 and pUB110 integrated into J3 regions. The system was validated with 99 MRSA strains carrying SCCmec elements of different types. The SCCmec types of 93 out of the 99 MRSA strains could be assigned. The SCCmec type assignments were identical to those made with a PCR system that uses numerous primer pairs to identify genes or gene alleles. Our system of six M-PCRs is thus a convenient and reliable method for typing SCCmec elements. Staphylococcal cassette chromosome mec (SCCmec) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for defining methicillin-resistant S. aureus (MRSA) clones in epidemiological studies. We have developed a convenient system for SCCmec type assignment. The system consists of six multiplex PCRs (M-PCRs) for identifying the ccr gene complex (ccr), the mec gene complex (mec), and specific structures in the junkyard (J) regions: M-PCR with primer set 1 (M-PCR 1) identified five types of ccr genes; M-PCR 2 identified class A to class C mec; M-PCRs 3 and 4 identified specific open reading frames in the J1 regions of type I and IV and of type II, III, and V SCCmec elements, respectively; M-PCR 5 identified the transposons Tn554 and ΨTn554 integrated into the J2 regions of type II and III SCCmec elements; and M-PCR 6 identified plasmids pT181 and pUB110 integrated into J3 regions. The system was validated with 99 MRSA strains carrying SCCmec elements of different types. The SCCmec types of 93 out of the 99 MRSA strains could be assigned. The SCCmec type assignments were identical to those made with a PCR system that uses numerous primer pairs to identify genes or gene alleles. Our system of six M-PCRs is thus a convenient and reliable method for typing SCCmec elements. Classifications Services AAC Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue AAC About AAC Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AAC RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0066-4804 Online ISSN: 1098-6596 Copyright © 2014 by the American Society for Microbiology. For an alternate route to AAC .asm.org, visit: AAC |
| Author | Teruyo Ito Keiichi Hiramatsu Shinya Watanabe Xiao Xue Ma Yoko Kondo Jerome Etienne Barry N. Kreiswirth |
| AuthorAffiliation | Juntendo University, Graduate School of Medicine, Department of Infection Control Science, Tokyo, Japan, 1 Juntendo University, Department of Bacteriology, Tokyo, Japan, 2 Public Health Research Institute, Newark, New Jersey, 3 Faculté de Médecine Laennec, Centre National de Référence des Staphylocoques, IFR62, INSERM E02030, 7 rue guillaume Paradin, 69008 Lyon, France 4 |
| AuthorAffiliation_xml | – name: Juntendo University, Graduate School of Medicine, Department of Infection Control Science, Tokyo, Japan, 1 Juntendo University, Department of Bacteriology, Tokyo, Japan, 2 Public Health Research Institute, Newark, New Jersey, 3 Faculté de Médecine Laennec, Centre National de Référence des Staphylocoques, IFR62, INSERM E02030, 7 rue guillaume Paradin, 69008 Lyon, France 4 |
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| Keywords | Genetic mapping Multiplex polymerase chain reaction Chromosome Identification CC chemokine Staphylococcus Infection Cassette Bacteriosis Bacteria Micrococcales Genetics Micrococcaceae Staphylococcal infection |
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| References | 15215121 - Antimicrob Agents Chemother. 2004 Jul;48(7):2637-51 12734443 - Curr Opin Infect Dis. 2003 Apr;16(2):103-24 3305073 - FEBS Lett. 1987 Aug 31;221(1):167-71 8665912 - Eur J Biochem. 1996 Mar 15;236(3):904-10 11897611 - Antimicrob Agents Chemother. 2002 Apr;46(4):1147-52 15712079 - Clin Infect Dis. 2005 Feb 15;40(4):562-73 8150070 - Eur J Epidemiol. 1993 Nov;9(6):658-62 6563036 - J Bacteriol. 1984 May;158(2):513-6 16000461 - J Clin Microbiol. 2005 Jul;43(7):3364-72 3878127 - Antimicrob Agents Chemother. 1985 Sep;28(3):397-403 10348769 - Antimicrob Agents Chemother. 1999 Jun;43(6):1449-58 11302791 - Antimicrob Agents Chemother. 2001 May;45(5):1323-36 12057957 - J Bacteriol. 2002 Jul;184(13):3623-9 3850810 - FEBS Lett. 1985 Nov 11;192(1):28-32 11408208 - Antimicrob Agents Chemother. 2001 Jul;45(7):1955-63 12654286 - Drug Resist Updat. 2003 Feb;6(1):41-52 15855533 - Antimicrob Agents Chemother. 2005 May;49(5):2070-83 18025671 - Methods Mol Biol. 2007;391:87-102 16081886 - J Clin Microbiol. 2005 Aug;43(8):3610-4 16126781 - J Antimicrob Chemother. 2005 Oct;56(4):624-32 12069968 - Antimicrob Agents Chemother. 2002 Jul;46(7):2155-61 16207957 - J Clin Microbiol. 2005 Oct;43(10):5026-33 16495263 - Antimicrob Agents Chemother. 2006 Mar;50(3):1001-12 10817707 - Antimicrob Agents Chemother. 2000 Jun;44(6):1549-55 12409412 - J Clin Microbiol. 2002 Nov;40(11):4289-94 11822775 - Microb Drug Resist. 2001 Winter;7(4):349-61 |
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Reddit... Staphylococcal cassette chromosome mec (SCCmec) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for... Staphylococcal cassette chromosome mec (SCC mec ) typing, in combination with genotyping of the Staphylococcus aureus chromosome, has become essential for... |
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| SubjectTerms | Anti-Bacterial Agents - pharmacology Antibiotics. Antiinfectious agents. Antiparasitic agents Bacterial diseases Bacterial Typing Techniques - methods Biological and medical sciences Chromosome Mapping Chromosomes, Bacterial - genetics DNA Transposable Elements DNA Transposable Elements - genetics DNA, Bacterial - chemistry DNA, Bacterial - genetics Human bacterial diseases Infectious diseases Mechanisms of Resistance Medical sciences Methicillin - pharmacology Methicillin Resistance - genetics Molecular Sequence Data Mutagenesis, Insertional Pharmacology. Drug treatments Polymerase Chain Reaction Polymerase Chain Reaction - methods Reproducibility of Results Sequence Analysis, DNA Staphylococcal infections, streptococcal infections, pneumococcal infections Staphylococcus Staphylococcus - classification Staphylococcus - drug effects Staphylococcus - genetics Staphylococcus aureus Staphylococcus aureus - classification Staphylococcus aureus - drug effects Staphylococcus aureus - genetics |
| Title | Combination of Multiplex PCRs for Staphylococcal Cassette Chromosome mec Type Assignment: Rapid Identification System for mec, ccr, and Major Differences in Junkyard Regions |
| URI | http://aac.asm.org/content/51/1/264.abstract https://www.ncbi.nlm.nih.gov/pubmed/17043114 https://journals.asm.org/doi/10.1128/AAC.00165-06 https://search.proquest.com/docview/19970123 https://pubmed.ncbi.nlm.nih.gov/PMC1797693 |
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