Research Article
Molecular Identification and Quantification of Tetracycline and Erythromycin Resistance Genes in Spanish and Italian Retail Cheeses
Table 1
Sequence and properties of the PCR primers used in this work.
| Application/primer | Target gene | Sequence (5′-3′) | Annealing (°C) | Amplicon size (bp) | Reference |
| Conventional PCR | | | | | | Tet-F | tet
a | GCTCA(T/C)GTTGA(T/C)GCAGGAA | 50 | 1292 |
[25] | Tet-R | AGGATTTGGCGG(C/G)ACTTC(G/T)A | TetK-F | tet(K) | TTATGGTGGTTGTAGCTAGAAA | 55 | 348 |
[15] | TetK-R | AAAGGGTTAGAAACTCTTGAAA | TetL-F | tet(L) | GTMGTTGCGCGCTATATTCC | 55 | 696 |
[15] | TetL-R | GTGAAMGRWAGCCCACCTAA | DI-F | tet(M) | GAYACICCIGGICAYRTIGAYTT | 55 | 1513 |
[26] | TetM-R | CACCGAGCAGGGATTTCTCCAC | TetO-F | tet(O) | AATGAAGATTCCGACAATTT | 55 | 781 |
[26] | TetO-R | CTCATGCGTTGTAGTATTCCA | TetS-F | tet(S) | ATCAAGATATTAAGGAC | 55 | 573 |
[26] | TetS-R | TTCTCTATGTGGTAATC | TetW-F | tet(W) | AAGCGGCAGTCACTTCCTTCC | 50 | 1150 |
[11] | Tet-R | AGGATTTGGCGG(C/G)ACTTC(G/T)A | ErmB-F | erm(B) | GAAAAGGTACTCAACCAAATA | 50 | 639 |
[27] | ErmB-R | AGTAACGGTACTTAAATTGTTTAC | ErmF-F | erm(F) | CGGGTCAGCACTTTACTATTG | 50 | 466 |
[27] | ErmF-R | GGACCTACCTCATAGACAAG | Real-time qPCR | | | | | | TBA-F | 16S rDNA | CGGCAACGAGCGCAACCC | 60 | 130 |
[28] | TBA-R | CCATTGTAGCACGTGTGTAGCC | TetK-qPCR-F | tet(K) | TGCTGCATTCCCTTCACTGA | 60 | 69 | This study | TetK-qPCR-R | GCTTTGCCTTGTTTTTTTCTTGTAA | TetL-qPCR-F | tet(L) | GGGTAAAGCATTTGGTCTTATTGG | 60 | 63 | This study | tetL-qPCR-R | ATCGCTGGACCGACTCCTT | TetM-qPCR-F | tet(M) | CAGAATTAGGAAGCGTGGACAA | 60 | 67 | This study | TetM-qPCR-R | CCTCTCTGACGTTCTAAAAGCGTAT | TetO-qPCR-F | tet(O) | AATGTCAGAACTGGAACAGGAAGAA | 60 | 59 | This study | TetO-qPCR-R | CGTGATAAACGGGAAATAACGTT | TetS-qPCR-F | tet(S) | CGAGGTCATTCTCATTGGTGAA | 60 | 84 | This study | TetS-qPCR-R | CAGACACTGCGTCCATTTGTAAA | TetW-qPCR-F | tet(W) | ACGGCAGCGCAAAGAGAA | 60 | 60 | This study | TetW-qPCR-R | CGGGTCAGTATCCGCAAGTT | ErmB-qPCR-F | erm(B) | GGATTCTACAAGCGTACCTTGGA | 60 | 69 | This study | ErmB-qPCR-R | AATCGAGACTTGAGTGTGCAAGAG | ErmF-qPCR-F | erm(F) | TGATGCCCGAAATGTTCAAGT | 60 | 63 | This study | ErmF-qPCR-R | AAAGGAAATTTCGGAACTGCAA | DGGE assays | | | | | | TetM-F | tet(M) | ACAGAAAGCTTATTATATAAC | 55 | 171 |
[15] | GCb-TetM-R | TGGCGTGTCTATGATGTTCAC | TetW-F | tet(W) | GAGAGCCTGCTATATGCCAGC | 64 | 168 |
[15] | GC-TetW-R | GGGCGTATCCACAATGTTAAC |
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tet, genes encoding ribosome protection proteins.
bSequence of the GC clamp: CGCCCGGGGCGCGCCCCGGGCGGGGGGGGGGCACGGGGGG.
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