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]
GC^b-TetM-R		TGGCGTGTCTATGATGTTCAC
TetW-F	tet(W)	GAGAGCCTGCTATATGCCAGC	64	168	[15]
GC-TetW-R		GGGCGTATCCACAATGTTAAC

tet, genes encoding ribosome protection proteins.
^bSequence of the GC clamp: CGCCCGGGGCGCGCCCCGGGCGGGGGGGGGGCACGGGGGG.