Table 4 Target genes, oligonucleotide primers, and PCR conditions used for the detection of virulence factors and antibiotic resistance genes in the S. epidermidis strains isolated from various types of hospital infectious samples
Target gene | Primer sequence (5′-3′) | PCR product (bp) | PCR programs | PCR volume (50 μL) |
|---|---|---|---|---|
vatA | F: TGGTCCCGGAACAACATTTAT R: TCCACCGACAATAGAATAGGG | 268 | 1 cycle: 94 °C, 5 min 30 cycles: 94 °C, 60 s 60 °C, 60 s 72 °C, 90 s 1 cycle: 72 °C, 7 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 |
vatB | F: GCTGCGAATTCAGTTGTTACA R: CTGACCAATCCCACCATTTTA | 136 | ||
vatC | F: AAGGCCCCAATCCAGAAGAA R: TCAACGTTCTTTGTCACAACC | 467 | ||
mecA | F: AAAATCGATGGTAAAGGTTGGC R: AGTTCTGCAGTACCGGATTTGC | 532 | ||
tetK | F: GTAGCGACAATAGGTAATAGT R: GTAGTGACAATAAACCTCCTA | 360 | ||
tetM | F: AGTGGAGCGATTACAGAA R: CATATGTCCTGGCGTGTCTA | 158 | 1 cycle: 94 °C, 5 min 30 cycles: 94 °C, 60 s 60 °C, 60 s 72 °C, 90 s 1 cycle: 72 °C, 7 | 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 |
msrA | F: GGCACAATAAGAGTGTTTAAAGG R: C AAGTTATATCATGAATAGATTGTCCTGTT | 940 | ||
msrB | F: TATGATATCCATAATAATTATCCAATC R: AAGTTATATCATGAATAGATTGTCCTGTT | 595 | ||
aacA-D | F: TAATCCAAGAGCAATAAGGGC R: GCCACACTATCATAACCACTA | 227 | ||
ermA | F: AAGCGGTAAACCCCTCTGA R: TTCGCAAATCCCTTCTCAAC | 190 | ||
ermC | F: AATCGTCAATTCCTGCATGT R: AATCGTCAATTCCTGCATGT | 229 | ||
linA | F: GGTGGCTGGGGGGTAGATGTATTAACTGG R: GCTTCTTTTGAAATACATGGTATTTTTCGA | 323 | ||
tsst-1 | F: ATGGCAGCATCAGCTTGATA R: TTTCCAATAACCACCCGTTT | 350 | 1 cycle: 94 °C, 6 min 30 cycles: 94 °C, 2 min 55 °C, 2 min 72 °C, 1 min 1 cycle: 72 °C, 8 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 |
etA | F: CTAGTGCATTTGTTATTCAA R: TGCATTGACACCATAGTACT | 119 | ||
etB | F: ACGGCTATATACATTCAATT R: TCCATCGATAATATACCTAA | 200 | ||
agrI | F: ATGCACATGGTGCACATGC R: GTCACAAGTACTATAAGCTGCGAT | 441 | 1 cycle: 94 °C, 6 min 26 cycle: 94 °C, 30 s 55 °C, 30 s 72 °C, 1 min 1 cycle: 72 °C, 8 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 |
agrII | F: ATGCACATGGTGCACATGC R: TATTACTAATTGAAAAGTGGCCATAGC | 575 | ||
agrIII | F: ATGCACATGGTGCACATGC R: GTAATGTAATAGCTTGTATAATAATACCCAG | 323 | ||
coa | F: CGAGACCAAGATTCAACAAG R: AAAGAAAACCACTCACATCA | 970 | 1 cycle: 95 °C, 2 min 30 cycles: 95 °C, 30 s 58 °C, 2 min 72 °C, 4 min 1 cycle: 72 °C, 7 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 |
clfA | F: GGCTTCAGTGCTTGTAGG R: TTTTCAGGGTCAATATAAGC | 980 | 1 cycle: 94 °C, 4 min 35 cycles: 94 °C, 1 min 57 °C, 1 min 72 °C, 1 min 1 cycle: 72 °C, 5 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 |
X-region | F: CAAGCACCAAAAGAGGAA R: CACCAGGTTTAACGACAT | 320 | 1 cycle: 95 °C, 4 min 25 cycles: 95 °C, 1 min 60 °C, 1 min 72 °C, 1 min 1 cycle: 72 °C, 3 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 |
IgG-binding region | F: CACCTGCTGCAAATGCTGCG R: GGCTTGTTGTTGTCTTCCTC | 920 | 1 cycle: 94 °C, 2 min 30 cycles: 94 °C, 1 min 58 °C, 1 min 72 °C, 1 min 1 cycle: 72 °C, 5 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 |