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, 34–46]
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 |