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Mechanisms of Resistance

Update to the Multiplex PCR Strategy for Assignment of mec Element Types in Staphylococcus aureus

Catarina Milheiriço, Duarte C. Oliveira, Hermínia de Lencastre
Catarina Milheiriço
1Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal
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Duarte C. Oliveira
1Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal
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  • For correspondence: dco@itqb.unl.pt
Hermínia de Lencastre
1Laboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal
2Laboratory of Microbiology, The Rockefeller University, New York, New York 10021
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DOI: 10.1128/AAC.00275-07
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    FIG. 1.

    Amplification patterns for the prototype strains obtained with the updated SCCmec multiplex PCR strategy. Lane 1, SCCmec type I (strain COL); lane 2, type II (N315); lane 3, type III (ANS46), lane 4, type IVa (MW2); lane 5, type IVb (8/6-3P); lane 6, type IVc (Q2314); lane 7, type IVd (JCSC4469); lane 8, type IVE (AR43/3330.1); lane 9, type IVg (M03-68); lane 10, type IVh (HAR22); lane 11, type V (WIS); lane 12, type VI (HDE288); lanes M, DNA molecular size marker (1-kb DNA Ladder Plus; Invitrogen Life Technologies, Carlsbad, CA).

Tables

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  • TABLE 1.

    Primers used in the updated version of SCCmec multiplex PCR

    Primer namePrimer sequence (5′→3′)Primer specificity (SCCmec type, region)Amplicon size (bp)Reference
    CIF2 F2TTCGAGTTGCTGATGAAGAAGGI, J1 region495 13
    CIF2 R2ATTTACCACAAGGACTACCAGC 13
    ccrC F2GTACTCGTTACAATGTTTGGV, ccr complex449This study
    ccrC R2ATAATGGCTTCATGCTTACCThis study
    RIF5 F10TTCTTAAGTACACGCTGAATCGIII, J3 region414 13
    RIF5 R13ATGGAGATGAATTACAAGGG 13
    SCCmec V J1 FTTCTCCATTCTTGTTCATCCV, J1 region377This study
    SCCmec V J1 RAGAGACTACTGACTTAAGTGGThis study
    dcs F2CATCCTATGATAGCTTGGTCI, II, IV, and VI, J3 region342 13
    dcs R1CTAAATCATAGCCATGACCG 13
    ccrB2 F2AGTTTCTCAGAATTCGAACGII and IV, ccr complex311This study
    ccrB2 R2CCGATATAGAAWGGGTTAGCThis study
    kdp F1AATCATCTGCCATTGGTGATGCII, J1 region284 13
    kdp R1CGAATGAAGTGAAAGAAAGTGG 13
    SCCmec III J1 FCATTTGTGAAACACAGTACGIII, J1 region243This study
    SCCmec III J1 RGTTATTGAGACTCCTAAAGCThis study
    mecI P2ATCAAGACTTGCATTCAGGCII and III, mec complex209 13
    mecI P3GCGGTTTCAATTCACTTGTC 13
    mecA P4TCCAGATTACAACTTCACCAGGInternal positive control162 13
    mecA P7CCACTTCATATCTTGTAACG 13
  • TABLE 2.

    Prototype strains and representative collection used for validation of updated SCCmec multiplex PCR assay

    StrainaOriginIsolation dateMLST (ST)bSCCmec typeccrAB typeMultiplex PCR assignmentReferencec
    COL United Kingdom1965250I1I 16
    UK13136United Kingdom1960250I1I 16
    BK793Egypt1961250I1I 16
    E2125Denmark1964247I1I 16
    PER34Spain1989250IA1I 16
    HPV107Portugal1992247IA1I 16
    BK1953United States1995247IA1I 16
    PER184Spain1991247IA1I 16
    PER88Spain1992247IA1I 16
    HAR40Belgium1995228I1I 15
    HAR21Finland20025I1I 15
    PL72Poland19915I-VARd1VI 15
    POL3Poland19925I-VAR1VI 15
    N315 Japan19825II2II 4
    USA100United States1995-20035II2II 10
    BK2464United States19965II2II 16
    JP1Japan19875II2II 16
    JP205Japan19985II2II 15
    USA200United States1995-200336II2II 10
    HAR24Finland200236II2II 15
    USA600United States1995-200345II2II 10
    ANS46 Australia1982239III3III 16
    R35United States1987239III3III 16
    HU25Brazil1993239III3III 16
    HUSA304Hungary1993239III3III 16
    HU106Hungary1996239III3III 16
    LHH1United States1994239III3III 16
    BK2421United States1996239III3III 16
    HSJ216Portugal1997239IIIA3III 16
    HDG2Portugal1993239IIIB3IIIB 16
    MW2 United States19981IVa2IV 1
    CA05 Chicago1999256IVa2IV 9
    8/6-3P Chicago1996770IVb2IV 9
    Q2314 Dallas, TX19965IVc2IV 6
    JCSC4469 Japan19825IVd2IVAB097677
    AR43/3330.1 Ireland1988-20028IVE2IV 17
    M03-68 Korea20035IVg2IV 8
    HAR22 Finland200222IVh2IV 11
    USA400United States1995-20031IV2IV 10
    USA800United States1995-20035IV2IV 10
    BM18United States19895IV2IV 16
    COB3Colombia19965IV2IV 16
    FFP311Portugal19965IV2IV 14
    VNG17Portugal1992-19935IV2IV 14
    RJP17Portugal1992-19935IV2IV 14
    HSA49Portugal19935IV2IV 15
    HSA74Portugal19935IV2IV 14
    ARG9Argentina19965IV2IV 14
    USA300United States1995-20038IV2IV 10
    USA500United States1995-20038IV2IV 10
    BK2529United States19968IV2IV 16
    BargII17United States19968IV2IV 16
    GRE120Greece19938IV2IV 15
    HAR36eGreece2002254IV-VAR2IV-VAR 15
    HGSA146Portugal200322IVh2IV 11
    HGSA163Portugal200322IVh2IV 11
    DEN2946Denmark200130IV2IV 15
    DEN2294Denmark200130IV2IV 15
    HAR38Belgium199545IV2IV 15
    USA700United States1995-200372IV2IV 10
    GRE14Greece199880IV2IV 15
    DEN2949Denmark200180IV2IV 15
    DEN114Denmark200180IV2IV 15
    DEN1451Denmark2001153IV2IV 15
    WIS AustraliaV ccrC V 5
    HT0087Australia2005377V ccrC V 18
    HT0184Greece2005377V ccrC V 18
    HT0350The Netherlands2003377V ccrC V 18
    HT0388Switzerland2003377V ccrC V 18
    HT0826France2003377V ccrC V 18
    HDE288 Portugal19965VI4VI 14
    HUC136Portugal19955VI4VI 14
    IPO92Portugal20015VI4VI 14
    • ↵ a Strains in boldface are the prototype strains for SCCmec types and subtypes.

    • ↵ b MLST, multilocus sequence type; ST, sequence type.

    • ↵ c Reference for SCCmec type assignment and ccr type.

    • ↵ d VAR, variant.

    • ↵ e Strain HAR36 is characterized by SCCmec type IV and is also positive for the J3 locus specific for SCCmec type III.

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Update to the Multiplex PCR Strategy for Assignment of mec Element Types in Staphylococcus aureus
Catarina Milheiriço, Duarte C. Oliveira, Hermínia de Lencastre
Antimicrobial Agents and Chemotherapy Aug 2007, 51 (9) 3374-3377; DOI: 10.1128/AAC.00275-07

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Update to the Multiplex PCR Strategy for Assignment of mec Element Types in Staphylococcus aureus
Catarina Milheiriço, Duarte C. Oliveira, Hermínia de Lencastre
Antimicrobial Agents and Chemotherapy Aug 2007, 51 (9) 3374-3377; DOI: 10.1128/AAC.00275-07
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KEYWORDS

Bacterial Proteins
methicillin resistance
Reverse Transcriptase Polymerase Chain Reaction
Staphylococcus aureus

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