Development of a melting-curve based multiplex real-time PCR assay for simultaneous detection of Streptococcus agalactiae and genes encoding resistance to macrolides and lincosamides

Table 2 Oligonucleotide primers of melting curve-based multiplex real-time PCR

Target^a	Nucleotide sequence (5′ to 3′)	Amplicon size (bp)	Reference
cfb ^b	F: CACACATGCTGTTGGAGTTCAGTTGA	138	This study
cfb ^b	R: ACGAAGTCGACAGCATCACACGAAA	138
erm(A)/(TR)	F: CCGGCAAGGAGAAGGTTATAATGA	190	Otaguiri et al. [5]
erm(A)/(TR)	R: GCATTCACCCGTTGACTCATTTCC	190
erm(B)	F: GCTCTTGCACACTCAAGTCTCGAT	117	Otaguiri et al. [5]
erm(B)	R: ACATCTGTGGTATGGCGGGTAAGT	117
mef(A/E)	F: GCGATGGTCTTGTCTATGGCTTCA	225	Otaguiri et al. [5]
mef(A/E)	R: AGCTGTTCCAATGCTACGGAT	225
RNaseP	F: AGATTTGGACCTGCGAGCG	64	WHO [27]
RNaseP	R: GAGCGGCTGTCTCCACAAGT	64
IGS1	F: GTCATTTCAGCTGGCGCCATCGATAC	260	Tavares et al. [32]
IGS1	R: TTGCCGCATAACGCATCTTAGCCA

^acfb gene encodes the CAMP factor; erm genes encode 23S rRNA methylases; mef gene encodes efflux pumps; RNaseP gene encodes human ribonuclease P; IGS1, intergenic spacer 1 of ribosomal RNA gene cluster of Cryptococcus gattii. ^bThe nucleotide sequences of Streptococcus agalactiae genes deposited in the GenBank/EMBL databases were used for specific primer design

ISSN: 1471-2393