Multiplex PCR for the simultaneous detection of the Enterobacterial gene wecA, the Shiga Toxin genes (stx1 and stx2) and the Intimin gene (eae)

Table 1 Primer sequences, product sizes and conditions used in the triplex and simplex PCRs

Primer sets	Target genes	Primers			MgCl₂ (mM)	Product size (bp)
Primer sets	Target genes	Names	Sequences (5′–3′)^a	μM	MgCl₂ (mM)	Product size (bp)
A	wecA	Meca202U	GGGTTGTCCTGCGTCTCGTT	0.05	3	452
	wecA	Meca633L	TATTCTGCCAGCACGCCAATG	0.05		452
	stx₁ and stx₂	UstxU1	TRTTGARCRAAATAATTTATATGT	0.5		526 (stx₁)
	stx₁ and stx₂	UstxL1	MTGATGATGRCAATTCAGTAT	0.5		523 (stx₂)
	stxA _2f	UstxU3	AATGGAACGGAATAACTTATATGT	0.05		523
	stxA _2f	UstxL3	GGTTGAGTGGCAATTAAGGAT	0.05		523
	eae intimin	Ueae28U	ACCCGGCACAAGCATAAG	0.1		741
	eae intimin	Ueae748L	CGTAAAGCGRGAGTCAATRTA	0.1		741
B	stx₁ and stx₂	UstxL1	MTGATGATGRCAATTCAGTAT	0.1	3
	stx ₁	Nestx1	GTACAACACTKGATGATCTC	0.3		200
	stx ₂	Nestx2	TGACRACGGACAGCAGT	0.05		410
C	wecA	Meca480UU21	GGATATGGTGGCGATTATGTA	0.2	2	226
		Meca685LU21	GAATGCTAGCAAAAAGAGCAC	0.2	2	226
		Meca479UU21	TGGATATGGTGGCGATTATGT	0.2	2	261
		Meca722LU18	TCCAGGCMCGCTTAATGC	0.2	2	261

Triplex PCR (A) semi-nested duplex PCR for stx₁ and stx₂ typing (B) and simplex PCR for Enterobacteriaceae (C)
^aR = A or G, M = A or C, K = G or T

ISSN: 1756-0500