Article Figures & Data
Figures
- 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
- TABLE 1.
Primers used in the updated version of SCCmec multiplex PCR
Primer name Primer sequence (5′→3′) Primer specificity (SCCmec type, region) Amplicon size (bp) Reference CIF2 F2 TTCGAGTTGCTGATGAAGAAGG I, J1 region 495 13 CIF2 R2 ATTTACCACAAGGACTACCAGC 13 ccrC F2 GTACTCGTTACAATGTTTGG V, ccr complex 449 This study ccrC R2 ATAATGGCTTCATGCTTACC This study RIF5 F10 TTCTTAAGTACACGCTGAATCG III, J3 region 414 13 RIF5 R13 ATGGAGATGAATTACAAGGG 13 SCCmec V J1 F TTCTCCATTCTTGTTCATCC V, J1 region 377 This study SCCmec V J1 R AGAGACTACTGACTTAAGTGG This study dcs F2 CATCCTATGATAGCTTGGTC I, II, IV, and VI, J3 region 342 13 dcs R1 CTAAATCATAGCCATGACCG 13 ccrB2 F2 AGTTTCTCAGAATTCGAACG II and IV, ccr complex 311 This study ccrB2 R2 CCGATATAGAAWGGGTTAGC This study kdp F1 AATCATCTGCCATTGGTGATGC II, J1 region 284 13 kdp R1 CGAATGAAGTGAAAGAAAGTGG 13 SCCmec III J1 F CATTTGTGAAACACAGTACG III, J1 region 243 This study SCCmec III J1 R GTTATTGAGACTCCTAAAGC This study mecI P2 ATCAAGACTTGCATTCAGGC II and III, mec complex 209 13 mecI P3 GCGGTTTCAATTCACTTGTC 13 mecA P4 TCCAGATTACAACTTCACCAGG Internal positive control 162 13 mecA P7 CCACTTCATATCTTGTAACG 13 - TABLE 2.
Prototype strains and representative collection used for validation of updated SCCmec multiplex PCR assay
Straina Origin Isolation date MLST (ST)b SCCmec type ccrAB type Multiplex PCR assignment Referencec COL United Kingdom 1965 250 I 1 I 16 UK13136 United Kingdom 1960 250 I 1 I 16 BK793 Egypt 1961 250 I 1 I 16 E2125 Denmark 1964 247 I 1 I 16 PER34 Spain 1989 250 IA 1 I 16 HPV107 Portugal 1992 247 IA 1 I 16 BK1953 United States 1995 247 IA 1 I 16 PER184 Spain 1991 247 IA 1 I 16 PER88 Spain 1992 247 IA 1 I 16 HAR40 Belgium 1995 228 I 1 I 15 HAR21 Finland 2002 5 I 1 I 15 PL72 Poland 1991 5 I-VARd 1 VI 15 POL3 Poland 1992 5 I-VAR 1 VI 15 N315 Japan 1982 5 II 2 II 4 USA100 United States 1995-2003 5 II 2 II 10 BK2464 United States 1996 5 II 2 II 16 JP1 Japan 1987 5 II 2 II 16 JP205 Japan 1998 5 II 2 II 15 USA200 United States 1995-2003 36 II 2 II 10 HAR24 Finland 2002 36 II 2 II 15 USA600 United States 1995-2003 45 II 2 II 10 ANS46 Australia 1982 239 III 3 III 16 R35 United States 1987 239 III 3 III 16 HU25 Brazil 1993 239 III 3 III 16 HUSA304 Hungary 1993 239 III 3 III 16 HU106 Hungary 1996 239 III 3 III 16 LHH1 United States 1994 239 III 3 III 16 BK2421 United States 1996 239 III 3 III 16 HSJ216 Portugal 1997 239 IIIA 3 III 16 HDG2 Portugal 1993 239 IIIB 3 IIIB 16 MW2 United States 1998 1 IVa 2 IV 1 CA05 Chicago 1999 256 IVa 2 IV 9 8/6-3P Chicago 1996 770 IVb 2 IV 9 Q2314 Dallas, TX 1996 5 IVc 2 IV 6 JCSC4469 Japan 1982 5 IVd 2 IV AB097677 AR43/3330.1 Ireland 1988-2002 8 IVE 2 IV 17 M03-68 Korea 2003 5 IVg 2 IV 8 HAR22 Finland 2002 22 IVh 2 IV 11 USA400 United States 1995-2003 1 IV 2 IV 10 USA800 United States 1995-2003 5 IV 2 IV 10 BM18 United States 1989 5 IV 2 IV 16 COB3 Colombia 1996 5 IV 2 IV 16 FFP311 Portugal 1996 5 IV 2 IV 14 VNG17 Portugal 1992-1993 5 IV 2 IV 14 RJP17 Portugal 1992-1993 5 IV 2 IV 14 HSA49 Portugal 1993 5 IV 2 IV 15 HSA74 Portugal 1993 5 IV 2 IV 14 ARG9 Argentina 1996 5 IV 2 IV 14 USA300 United States 1995-2003 8 IV 2 IV 10 USA500 United States 1995-2003 8 IV 2 IV 10 BK2529 United States 1996 8 IV 2 IV 16 BargII17 United States 1996 8 IV 2 IV 16 GRE120 Greece 1993 8 IV 2 IV 15 HAR36e Greece 2002 254 IV-VAR 2 IV-VAR 15 HGSA146 Portugal 2003 22 IVh 2 IV 11 HGSA163 Portugal 2003 22 IVh 2 IV 11 DEN2946 Denmark 2001 30 IV 2 IV 15 DEN2294 Denmark 2001 30 IV 2 IV 15 HAR38 Belgium 1995 45 IV 2 IV 15 USA700 United States 1995-2003 72 IV 2 IV 10 GRE14 Greece 1998 80 IV 2 IV 15 DEN2949 Denmark 2001 80 IV 2 IV 15 DEN114 Denmark 2001 80 IV 2 IV 15 DEN1451 Denmark 2001 153 IV 2 IV 15 WIS Australia V ccrC V 5 HT0087 Australia 2005 377 V ccrC V 18 HT0184 Greece 2005 377 V ccrC V 18 HT0350 The Netherlands 2003 377 V ccrC V 18 HT0388 Switzerland 2003 377 V ccrC V 18 HT0826 France 2003 377 V ccrC V 18 HDE288 Portugal 1996 5 VI 4 VI 14 HUC136 Portugal 1995 5 VI 4 VI 14 IPO92 Portugal 2001 5 VI 4 VI 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.
