Table 4.

Detection of virulence genes by multiplex PCR^a

	Gene (amplicon bp)	Annealing condition	Reference for multiplex composition
Multiplex 1	afaBCIII (750) papC (328) sfaDE (410)	65 °C – 1 min	[47]
Multiplex 2	cvaC (680) KpsMT II (272) ompT (559)	58 °C – 30 s	This study
Multiplex 3	hlyA (1,177) iroN (665) papA (720) traT (290)	58 °C – 30 s	This study
Multiplex 4	ompA (919) hra (537)	58 °C – 30 s	This study
Multiplex 5	iha (925) sat (667) sitA (608)	58 °C – 30 sec	This study

^aprimers for multiplex PCR reactions 2, 3, 4, and 5 were described elsewhere [46]. Genes chuA, iucD,irp2, vat, and fimA were searched by single PCR reactions [46]