시험법
특성 | Genes | Primer | Sequence | Anealing T (℃) | Amplicon |
---|---|---|---|---|---|
내성 유전자 | mecA1 | mA1 | TGCTATCCACCCTCAAACAGG | 59 | 286 |
mA2 | AACGTTGTAACCACCCCAAGA | ||||
mecC2 | mecC MFP | GAAAAAAAGGCTTAGAACGCCTC | 60 | 138 | |
mecC MRP | GAAGATCTTTTCCGTTTTCAGC | ||||
분자역학 | SCCmec3 | ccrA1 | CAAGCCTTATCAGGTACGAA | 56 |
5: 202 bp 2: 307 bp 4: 406 bp 7: 519 bp 1: 672 bp 8: 802 bp 3: 982 bp |
ccrA2 | CATTACGTCAACAACCGCAA | ||||
ccrA3 | CTGAATCATTCAGAAAACAGAC | ||||
ccrA4 | GTCCTAAAACAGCAACAAATGA | ||||
ccrB1 | GAGCATTAACTTGCCTGTTG | ||||
ccrB2 | CCTTCTGTGCTTTGCATTTC | ||||
ccrB3 | GACCTTCGTTCGCATCTTTT | ||||
ccrB4 | GGTTACAGTATTCAAGGTCAAT | ||||
ccrB6 | CTGGTATGTTATTATATCCTAAAG | ||||
ccrC1 | GCAATGAAACGTCTATTACAAG | ||||
ccrC2 | CAAACATTGTAACGAGTACTTC | ||||
mecA_univ | CTGCTATCTTTATAAACTTGTTTG | 56 | A: 1358 bp B: 1235 bp C1: 717 bp C2: 533 bp E: 979 bp |
||
mecA_E | ACATAACCTAAAAGGTGTACTG | ||||
mecR1_B | TCATGTGAAGCTCGATATACTA | ||||
ecR1_E | ACCAAACTTTATGATTTAACTGAG | ||||
merR1_A | TTCATTATAAAGCACAAAACTTCC | ||||
IS431_C2 | GTTGAATGATGAACGTTTACAC | ||||
IS431_C1 | GGGACATACATTAGATATTTGG | ||||
1. PCR 조건: 94℃ 2 min + 30X (94℃ 2 min + 57℃ 1 min + 72℃ 2 min) + 72℃ 2 min 2. PCR 조건: 94℃ 15 min + 30X (94℃ 30sec + 59℃ 1 min + 72℃ 1 min) + 72℃ 10 min 3. ccr gene PCR 조건: 95℃ 5 min + 28X (97℃ 10sec + 56℃ 20sec + 72℃ 40 sec) + 72℃ 7 min 4. mec gene PCR 조건: 95℃ 5 min + 28X (97℃ 10sec + 56℃ 20sec + 72℃ 1 min) + 72℃ 7 min |
SCCmec type | ccr (bp) | mec gene complex (bp) |
---|---|---|
I | 672 | 1235 |
II | 307 | 1358 |
III | 982 | 1358 |
IV | 307 | 1235 |
V | 202 | 533 |
VI | 406 | 1235 |
VII | 202 | 717 |
VIII | 406 | 1358 |
IX | 672 | 533 |
X | 517 | 717 |
XI | 802 | 979 |
Target genes | 참조균주 | |
---|---|---|
내성유전자 | mecA | SA ROH0001G |
SCCmec | SCCmec I | ATCC BAA-44 |
SCCmec II | SA ROH0028G | |
SCCmec III | ATCC 33592 | |
SCCmec IV | SA ROH0029G | |
SCCmec V | ATCC BAA-2094 | |
SCCmec VII | SA ROH0030G | |
SCCmec XI (mecC) | ATCC BAA-2313 | |
Typing | agr I | SA ROH0004G |
agr II | SA ROH0005G | |
agr III | SA ROH0006G | |
agrIV | SA ROH0007G |
항목 | 유전자 | Primer | Sequence | Anealing T (℃) | Amplicon |
---|---|---|---|---|---|
내성 유전자 | vanA1 | vanA F | AGCTGTACTCTCGCCGGATA | 52 | 320 |
vanAR | CCGGCTTAACAAAAACAGGA | ||||
vanB1 | vanB F | GGCTCAGAAAATGCGATGAT | 52 | 250 | |
vanB R | TGTCAATCAGTGCAGGAAGC | ||||
vanM2 | vanHM F | CAGCGTGGGGCACAAGTCTGA | 58 | 377 | |
vanHM R | TGCCGTACGCCAACACGTGA | ||||
1. PCR 조건: 94℃ 3 min + 30X (94℃ 1 min + 52℃ 30sec + 72℃ 30sec) + 72℃ 10 min 2. PCR 조건: 94℃ 3 min + 30X (94℃ 1 min + 55℃ 30sec + 72℃ 30sec) + 72℃ 10 min |
Target genes | 참조균주 | |
---|---|---|
내성유전자 | vanA | EM ROH0015G |
vanB | ATCC 51575 |
항목 | Target gene | Primer name | Primer sequence | Anealing T (℃) | Amplicon size (bp) |
---|---|---|---|---|---|
내성 유전자 | blaOXA-231 | OXA-23 F | CTCTTTTTCTTTCTGGTTGTAC | 54 | 776 |
OXA-23 R | CAGCTGTTTTAATGATTTCATC | ||||
blaOXA-241 | OXA-24 F | CTTCCTATATTCAGCATTTCTATT | 56 | 807 | |
OXA-24 R | GATTCCAAGATTTTCTAGCGAC | ||||
blaOXA-581 | OXA-58 F | GCTTAAGCATAAGTATTGGGG | 55 | 793 | |
OXA-58 R | CCCAACTTATCTAGCACATC | ||||
blaOXA-481 | OXA-48 F | TTGGTGGCATCGATTATCGG | 57 | 744 | |
OXA-48 R | GAGCACTTCTTTTGTGATGGC | ||||
blaOXA-1432 | OXA-143 F | CTTCCTATTCTCAGCATTTCTAC | 56 | 820 | |
OXA-143 R | CCCTAAATTATATAATCCCTAAATTC | ||||
blaOXA-2352 | OXA-235 F | GAAAACTCTTATTTTGTTGCCTTTA | 56 | 826 | |
OXA-235 R | CCCTTCAGCTTTCGGATAAAG | ||||
blaOXA-511 | OXA-51 F | GAACATTAAAACACTCTTACTTAT | 56 | 823 | |
OXA-51 R | TTAGAACAATTAGGTATTTTATAG | ||||
ISAba1-blaOXA-512 | ISAba1-OXA-51 F | CCCATTTCCAATTGGTTCTATC | 56 | 570 | |
ISAba1-OXA-51 R | CCATAGCTTTGTTGAGTTTGG | ||||
blaTEM3 | TEM F | ATGAGTATTCAACATTTCCGT | 59 | 861 | |
TEM R | TTACCAATGCTTAATCAGTGA | ||||
blaSHV3 | SHV F | CCGGGTTATTCTTATTTGTCGCT | 61 | 927 | |
SHV R | TAGCGTTGCCAGTGCTCG | ||||
blaCTX-M-11 | CTXM-1 F | ACCGTCACGCTGTTGTTAGG | 56 | 819 | |
CTXM-1 R | GTCGGTGACGATTTTAGCCG | ||||
blaCTX-M-21 | CTXM-2 F | AATGTTAACGGTGATGGCGA | 56 | 844 | |
CTXM-2 R | ACCGTGGGTTACGATTTTCG | ||||
blaCTX-M-91 | CTXM-9 F | GTGCAACGGATGATGTTCG | 56 | 845 | |
CTXM-9 R | ATGATTCTCGCCGCTGAAG | ||||
blaCTX-M-251 | CTXM-25 F | GTAAGGCGGGCGATGTTAAT | 56 | 856 | |
CTXM-25 R | AACCGTCGGTGACAATTCTG | ||||
blaKPC2 | KPC F | ATGTCACTGTATCGCCGTCT | 56 | 893 | |
KPC R | TTTTCAGAGCCTTACTGCCC | ||||
blaPER4 | PER F | GAATGTCATTATAAAAGCTGTAGT | 55 | 915 | |
PER R | CTTAGGGCAGAAAGCTTTTTC | ||||
blaGES2 | GES F | GCGCTTCATTCACGCACTAT | 56 | 753 | |
GES R | GCGTAATCTCTCTCCTGGGC | ||||
blaIMP2 | IMP F | TGAGCAATGTATCTGTATTC | 56 | 740 | |
IMP R | TTAGTTGCTTGGTTTTGATG | ||||
blaVIM2 | VIM F | TGGTCTACATGACCGCGTCT | 56 | 766 | |
VIM R | CGACTGAGCGATTTGTGTG | ||||
blaNDM-12 | NDM-1 F | CAATATTATGCACCCGGTCG | 56 | 726 | |
NDM-1 R | ATCATGCTGGCCTTGGGGAA | ||||
blaCMY-15 | MOXM F | CAACAACGACAATCCATCCTG | 58 | 1086 | |
MOXM R | GTTGCGATTGGCCAGCATG | ||||
blaCMY-25 | CIT F | TGATGAAAAAATCGTTATGCTCC | 56 | 1097 | |
CIT R | GTTAGGATAGCTTTTGTTTGCC | ||||
blaDHA6 | DHA F | GTTATCTGCAACACTGATTTCC | 56 | 1118 | |
DHA R | CACTCAAAATAGCCTGTGCAG | ||||
blaACC6 | ACC F | GAACACATTGAAGCTGTTATCC | 56 | 1153 | |
ACC R | CTTATTCCCTTCCAATGAGCTC | ||||
blaACT-16 | ACT F | GATGACAAAATCCCTAAGCTGT | 56 | 1098 | |
ACT R | GGGTTCGGATAGCTTTTATTC | ||||
blaFOX6 | FOX F | CATTATCCAGCCGATGCTCAA | 58 | 1001 | |
FOX R | TGGCCTCGATGGGATAGTTG | ||||
mcr-17 | CLR5-F | CGGTCAGTCCGTTTGTTC | 56 | 309 | |
CLR5-R | CTTGGTCGGTCTGTAGGG | ||||
mcr-1 / -28 | mcr-12-281F | CTTATGGCACGGTCTATGA | 54 | 650 | |
mcr-12-930R | CACATTTTCTTGGTATTTGG | ||||
1. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 40sec) + 72℃ 7 min 2. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 30sec) + 72℃ 7 min 3. PCR 조건: 94℃ 5 min + 35X (94℃ 30sec + Anealing T(℃) 30sec + 72℃ 30sec) + 72℃ 7 min 4. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 1 min) + 72℃ 7 min 5. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + Anealing T(℃) 30sec + 72℃ 1 min) + 72℃ 7 min 6. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 1 min) + 72℃ 7 min 7. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 56℃ 20sec + 72℃ 20sec) + 72℃ 7 min 8. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 54℃ 20sec + 72℃ 40sec) + 72℃ 7 min |
Target genes | 참조균주 | 보유유전자 | |
---|---|---|---|
내성유전자 | CTX-M1 | EC ROH0016G | CTX-M15 |
CTX-M1 | EC ROH0031G | CTX-M55 | |
CTX-M1 | EC ROH0032G | CTX-M186 | |
CTX-M9 | EC ROH0017G | CTX-M14 | |
CTX-M9 | EC ROH0033G | CTX-M27 | |
TEM | EC ROH0018G | TEM-1 | |
SHV | KP ROH0019G | SHV-1 | |
IMP | PA ROH0020G | IMP-6 | |
VIM | PA ROH0021G | VIM-2 | |
GES | KP ROH0022G | GES-5 | |
NDM | EC ROH0023G | NDM-1 | |
NDM | EC ROH0034G | NDM-5 | |
KPC | KP ROH0024G | KPC-2 | |
OXA-48 | KP ROH0025G | OXA-232 | |
DHA-1 | EC ROH0026G | DHA-1 | |
CMY-2 | EC ROH0027G | CMY-2 |
Carbapenem에 대해서 중간이거나 내성인 균주 및 BMD로 시행한 감수성 시험에서 MIC ≥ 4 μg/mL
항목 | Target gene | Primer name | Primer sequence | Anealing T (℃) | Amplicon size (bp) |
---|---|---|---|---|---|
내성 유전자 | bla PER1 | PER F | GAATGTCATTATAAAAGCTGTAGT | 55 | 915 |
PER R | CTTAGGGCAGAAAGCTTTTTC | ||||
bla GES2 | GES F | GCGCTTCATTCACGCACTAT | 56 | 753 | |
GES R | GCGTAATCTCTCTCCTGGGC | ||||
bla IMP2 | IMP F | TGAGCAATGTATCTGTATTC | 56 | 740 | |
IMP R | TTAGTTGCTTGGTTTTGATG | ||||
bla VIM2 | VIM F | TGGTCTACATGACCGCGTCT | 56 | 766 | |
VIM R | CGACTGAGCGATTTGTGTG | ||||
bla NDM-12 | NDM-1 F | CAATATTATGCACCCGGTCG | 56 | 726 | |
NDM-1 R | ATCATGCTGGCCTTGGGGAA | ||||
mcr -13 | CLR5-F | CGGTCAGTCCGTTTGTTC | 56 | 309 | |
CLR5-R | CTTGGTCGGTCTGTAGGG | ||||
mcr -1 / -24 | mcr-12-281F | CTTATGGCACGGTCTATGA | 54 | 650 | |
mcr-12-930R | CACATTTTCTTGGTATTTGG | ||||
1. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + 55℃ 20sec + 72℃ 1 min) + 72℃ 7 min 2. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 30sec) + 72℃ 7 min 3. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 56℃ 20sec + 72℃ 20sec) + 72℃ 7 min 4. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 54℃ 20sec + 72℃ 40sec) + 72℃ 7 min |
Target genes | 참조균주 | 보유유전자 | |
---|---|---|---|
내성유전자 | IMP | PA ROH0020G | IMP-6 |
VIM | PA ROH0021G | VIM-2 | |
GES | KP ROH0022G | GES-5 | |
NDM | EC ROH0023G | NDM-1 | |
NDM | EC ROH0034G | NDM-5 |
Carbapenem에 대해서 중간이거나 내성인 균주 및 BMD로 시행한 감수성 시험에서 MIC ≥ 4 μg/mL일 때 시행
항목 | Target gene | Primer name | Primer sequence | Anealing T (℃) | Amplicon size (bp) |
---|---|---|---|---|---|
내성 유전자 | blaOXA-231 | OXA-23 F | CTCTTTTTCTTTCTGGTTGTAC | 54 | 776 |
OXA-23 R | CAGCTGTTTTAATGATTTCATC | ||||
blaOXA-241 | OXA-24 F | CTTCCTATATTCAGCATTTCTATT | 56 | 807 | |
OXA-24 R | GATTCCAAGATTTTCTAGCGAC | ||||
blaOXA-581 | OXA-58 F | GCTTAAGCATAAGTATTGGGG | 55 | 793 | |
OXA-58 R | CCCAACTTATCTAGCACATC | ||||
blaOXA-481 | OXA-48 F | TTGGTGGCATCGATTATCGG | 57 | 744 | |
OXA-48 R | GAGCACTTCTTTTGTGATGGC | ||||
blaOXA-511 | OXA-51 F | GAACATTAAAACACTCTTACTTAT | 56 | 823 | |
OXA-51 R | TTAGAACAATTAGGTATTTTATAG | ||||
ISAba1-blaOXA-512 | ISAba1-OXA-51 F | CCCATTTCCAATTGGTTCTATC | 56 | 570 | |
ISAba1-OXA-51 R | CCATAGCTTTGTTGAGTTTGG | ||||
blaIMP2 | IMP F | TGAGCAATGTATCTGTATTC | 56 | 740 | |
IMP R | TTAGTTGCTTGGTTTTGATG | ||||
blaVIM2 | VIM F | TGGTCTACATGACCGCGTCT | 56 | 766 | |
VIM R | CGACTGAGCGATTTGTGTG | ||||
blaNDM-12 | NDM-1 F | CAATATTATGCACCCGGTCG | 56 | 726 | |
NDM-1 R | ATCATGCTGGCCTTGGGGAA | ||||
mcr-13 | CLR5-F | CGGTCAGTCCGTTTGTTC | 56 | 309 | |
CLR5-R | CTTGGTCGGTCTGTAGGG | ||||
mcr-1 / -24 | mcr-12-281F | CTTATGGCACGGTCTATGA | 54 | 650 | |
mcr-12-930R | CACATTTTCTTGGTATTTGG | ||||
1. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 40sec) + 72℃ 7 min 2. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 30sec) + 72℃ 7 min 3. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 56℃ 20sec + 72℃ 20sec) + 72℃ 7 min 4. PCR 조건: 94℃ 5 min + 25X (94℃ 30sec + 54℃ 20sec + 72℃ 40sec) + 72℃ 7 min |
Target genes | 참조균주 | 보유유전자 | |
---|---|---|---|
내성유전자 | OXA-23 | AB ROH0035G | OXA-23 |
IMP | PA ROH0020G | IMP-6 | |
VIM |
PA ROH0021G | VIM-2 | |
GES |
KP ROH0022G | GES-5 | |
NDM |
EC ROH0023G | NDM-1 | |
NDM | EC ROH0034G | NDM-5 |
Carbapenem에 대해서 중간이거나 내성인 균주 및 BMD로 시행한 감수성 시험에서 MIC ≥ 4 μg/mL일 때 시행
항목 | Target gene | Primer name | Primer sequence | Anealing T (℃) | Amplicon size (bp) |
---|---|---|---|---|---|
내성 유전자 |
qnrA1 | For | GGCCATGGATATTATTGATAA | 50 | 606 |
Rev | GGATCCGGGCAGCACTATTACTCC | ||||
qnrB12 | For | CAGATTTCCGCGGCGCAAGC | 60 | 353 | |
Rev | CGCGATGCCAAGTCGCTCCA | ||||
qnrB42 | For | TGACTCTGGCGTTAGTTGGCGA | 60 | 609 | |
Rev | GCAACGATGCCTGGTAGCTGTCC | ||||
qnrS2 | For | TGCTGCGTCAGGAACGGCTG | 60 | 466 | |
Rev | TGAGCGTGGGTCTCGCGGTA | ||||
aac(6ʼ)Ib-cr3 | For | ACGATTCCGTCACACTGCGCC | 60 | 524 | |
Rev | TCGCTCGAATGCCTGGCGTG | ||||
ermA4 | For | ACGATATTCACGGTTTACCCACTTA | 54 | 610 | |
Rev | AACCAGAAAAACCCTAAAGACACG | ||||
ermB4 | For | TAACGACGAAACTGGCTAAAAT | 54 | 416 | |
Rev | ATCTGTGGTATGGCGGGTAAG | ||||
1. PCR 조건: 95℃ 15 min + 40X (94℃ 1 min + 50℃ 2 min + 72℃ 3 min) + 72℃ 10 min 2. PCR 조건: 94℃ 5 min + 30X (94℃ 30sec + Anealing T(℃) 20sec + 72℃ 40sec) + 72℃ 7 min 3. PCR 조건: 94℃ 5 min + 30X (94℃ 45sec + 55℃ 45sec + 72℃ 60sec) + 72℃ 10 min 4. PCR 조건: 95℃ 15 min + 40X (94℃ 20sec + 53℃ 20sec + 72℃ 40sec) + 72℃ 10 min |