Multiplex Fast Real-Time PCR for Quantitative Detection and Identification of cos- and pac-Type Streptococcus thermophilus Bacteriophages

DOI: 10.1128/AEM.00295-08

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FIG. 1.
Real-time qPCR analysis of 10-fold serial dilutions in skim milk of (A) plasmid pEM212 DNA digested with BglII and bacteriophage φP13.2 and (B) plasmid pEM213 DNA digested with BglII and bacteriophage φipla124. C_T values were plotted against the logarithm of the calculated plasmid copy number or the number of PFU included in each reaction mixture.

TABLE 1.

PCR primers and TaqMan MGB probes used in this study

Specificity	Primer or probe	Sequence (5′-3′)^a	Position (bases)
orf1510 ^b	mgbPac2	VIC-ACATGGCTGCATCTCT-MGB (P)	13105-13120
	qPac1	CGGGTGCTGGTTTCAATCA (F)	13041-13060
	qPac2	CTGCTGAGTTATCACTAATCGAACC (R)	13143-13167
orf18 ^c	mgbCos	FAM-TTGGTCGTTCTACTGTTAA-MGB (P)	15874-15892
	qCos1	TGCCATATCATGTTGAGATAAGGAC (F)	15820-15844
	qCos2	TGCATCAACAATTTTATCGCCTTG (R)	15906-15929
pEM125	mgbIC	NED-CAAGCTCGAAATTAACCCTCACTAA-MGB (P)
	IC-FW	GAGTAGGTCATTTAAGTTGAGCATAATAGG (F)
	IC-R	CAAGCTCGAAATTAACCCTCACTAA (R)

↵ a (F), forward primer; (R), reverse primer; (P), TaqMan MGB probe.
↵ b orf1510 is the putative minor tail protein gene of the Sfi11 bacteriophage (accession no. AF158600).
↵ c orf18 is the antireceptor gene of the Sf21 bacteriophage (accession no. AF115103).