Table 1 Oligonucleotide primers and PCR conditions used for amplification of the antibiotic resistance genes and virulence factors in the A. baumannii strains of animal origin [19, 34,35,36,37,38,39,40,41,42,43,44,45,46]
| Target gene | Primer Sequence (5′-3′) | Size of product (bp) | PCR conditions | Volume (50 μl) |
|---|---|---|---|---|
| draBC |
GCTGGGCAGCAAACTGATAACTCTC CATCAAGCTGTTTGTTCGTCCGCCG | 750 |
1 cycle: 95 0C ------------ 4 min. 30 cycle: 95 0C ------------ 50 s 58 0C ------------ 60 s 72 0C ------------ 45 s 1 cycle: 72 0C ------------ 8 min |
5 μL PCR buffer 10X 1.5 mM Mgcl2 200 μM dNTP (Fermentas) 0.5 μM of each primers F & R 1.25 U Taq DNA polymerase (Fermentas) 2.5 μL DNA template |
| cnf1 |
AAGATGGAGTTTCCTATGCAGGAG CATTCAGAGTCCTGCCCTCATTATT | 498 | ||
| csgA |
ACTCTGACTTGACTATTACC AGATGCAGTCTGGTCAAC | 200 | ||
| cvaC |
CACACACAAACGGGAGCTGTT CTTCCCGCAGCATAGTTCCAT | 680 | ||
| iutA |
GGCTGGACATCATGGGAACTGG CGTCGGGAACGGGTAGAATCG | 300 | ||
| fyuA |
TGATTAACCCCGCGACGGGAA CGCAGTAGGCACGATGTTGTA | 880 | ||
| cnf2 |
AATCTAATTAAAGAGAAC CATGCTTTGTATATCTA | 543 |
1 cycle: 94 0C ------------ 6 min. 34 cycle: 95 0C ------------ 50 s 58 0C ------------ 70 s 72 0C ------------ 55 s 1 cycle: 72 0C ------------ 10 min |
5 μL PCR buffer 10X 2 mM Mgcl2 150 μM dNTP (Fermentas) 0.75 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 3 μL DNA template |
| kpsMT II |
GCGCATTTGCTGATACTGTTG CATCCAGACGATAAGCATGAGCA | 272 | ||
| PAI |
GGACATCCTGTTACAGCGCGCA TCGCCACCAATCACAGCCGAAC | 930 | ||
| papC |
GACGGCTGTACTGCAGGGTGTGGCG ATATCCTTTCTGCAGGGATGCAATA | 328 | ||
| fimH |
TGCAGAACGGATAAGCCGTGG GCAGTCACCTGCCCTCCGGTA | 508 |
1 cycle: 95 0C ------------ 4 min. 34 cycle: 94 0C ------------ 60 s 56 0C ------------ 45 s 72 0C ------------ 60 s 1 cycle: 72 0C ------------ 10 min |
5 μL PCR buffer 10X 2 mM Mgcl2 200 μM dNTP (Fermentas) 0.5 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 5 μL DNA template |
| ibeA |
AGGCAGGTGTGCGCCGCGTAC TGGTGCTCCGGCAAACCATGC | 170 | ||
| PapG II-III |
CTGTAATTACGGAAGTGATTTCTG ACTATCCGGCTCCGGATAAACCAT | 1070 | ||
| sfa/focDE |
CTCCGGAGAACTGGGTGCATCTTAC CGGAGGAGTAATTACAAACCTGGCA | 410 | ||
| traT |
GGTGTGGTGCGATGAGCACAG CACGGTTCAGCCATCCCTGAG | 290 | ||
| aadA1 |
TATCCAGCTAAGCGCGAACT ATTTGCCGACTACCTTGGTC | 447 |
1 cycle: 94 0C ------------ 6 min. 33 cycle: 95 0C ------------ 70 s 55 0C ------------ 65 s 72 0C ------------ 90 s 1 cycle: 72 0C ------------ 8 min |
5 μL PCR buffer 10X 2 mM Mgcl2 150 μM dNTP (Fermentas) 0. 5 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 2 μL DNA template |
| aac(3)-IV |
CTTCAGGATGGCAAGTTGGT TCATCTCGTTCTCCGCTCAT | 286 | ||
| sul1 |
TTCGGCATTCTGAATCTCAC ATGATCTAACCCTCGGTCTC | 822 | ||
| blaSHV |
TCGCCTGTGTATTATCTCCC CGCAGATAAATCACCACAATG | 768 | ||
| CITM |
TGGCCAGAACTGACAGGCAAA TTTCTCCTGAACGTGGCTGGC | 462 | ||
| cat1 |
AGTTGCTCAATGTACCTATAACC TTGTAATTCATTAAGCATTCTGCC | 547 | ||
| cmlA |
CCGCCACGGTGTTGTTGTTATC CACCTTGCCTGCCCATCATTAG | 698 | ||
| tet(A) |
GGTTCACTCGAACGACGTCA CTGTCCGACAAGTTGCATGA | 577 | ||
| tet(B) |
CCTCAGCTTCTCAACGCGTG GCACCTTGCTGATGACTCTT | 634 | ||
| dfrA1 |
GGAGTGCCAAAGGTGAACAGC GAGGCGAAGTCTTGGGTAAAAAC | 367 | ||
| qnr |
GGGTATGGATATTATTGATAAAG CTAATCCGGCAGCACTATTTA | 670 | ||
| imp |
GAATAGAATGGTTAACTCTC CCAAACCACTAGGTTATC | 188 |
1 cycle: 95 0C ------------ 4 min. 30 cycle: 95 0C ------------ 45 s 58 0C ------------ 60s 72 0C ------------ 40 s 1 cycle: 72 0C ------------ 5 min |
5 μL PCR buffer 10X 1.5 mM Mgcl2 100 μM dNTP (Fermentas) 1 μM of each primers F & R 1 U Taq DNA polymerase (Fermentas) 2.5 μL DNA template |
| vim |
GTTTGGTCGCATATCGCAAC AATGCGCAGCACCAGGATAG | 382 | ||
| sim |
GTACAAGGGATTCGGCATCG GTACAAGGGATTCGGCATCG | 569 | ||
| Oxa-23-like |
GATCGGATTGGAGAACCAGA ATTTCTGACCGCATTTCCAT | 501 |
1 cycle: 94 0C ------------ 5 min. 32 cycle: 95 0C ------------ 50 s 60 0C ------------ 60 s 72 0C ------------ 70 s 1 cycle: 72 0C ------------ 10 min |
5 μL PCR buffer 10X 2.5 mM Mgcl2 200 μM dNTP (Fermentas) 0.5 μM of each primers F & R 1.5 U Taq DNA polymerase (Fermentas) 2 μL DNA template |
| Oxa-24-like |
GGTTAGTTGGCCCCCTTAAA AGTTGAGCGAAAAGGGGATT | 246 | ||
| Oxa-51-like |
TAATGCTTTGATCGGCCTTG TGGATTGCACTTCATCTTGG | 353 | ||
| Oxa-58-like |
AAGTATTGGGGCTTGTGCTG CCCCTCTGCGCTCTACATAC | 599 |