Prevalence of E. coli ST131 among Uropathogenic E. coli Isolates from Iraqi Patients in Wasit Province, Iraq

Table 2

Components of 50 μl PCR master mix (2 pools) and amplification conditions for the surveillance of E. coli ST131 clone by conventional PCR assay.

Pool no.	Sterile DW (μl)	Primers	DNA (μl)	Amplification conditions
1 (three genes)	39	6 μl: 1 μl each of; (i) trpAST131-O16. f; (ii) trpAST131-O16. r; (iii) pabBST131-O25. f; (iv) pabBST131-O25. r; (v) uidA.f; (vi) uidA.r	5	(1) Initial denaturation for 4 min at 94°C; (2) 30 cycles of; (i) Denaturation for 5 s at 94°C; (ii) Annealing for 20 s at 63°C; (iii) Extension for 30 s at 72°C; (3) Final extension at 72°C for 5 min
2 (three genes)	39	6 μl: (i) 2 μ1 gndbis.f; (ii) 1 μl rfbO16.r; (iii) 1 μl rfbO25b.r; (iv) 1 μl uidA.f; (v) 1 μl uidA.r	5	(1) Initial denaturation for 4 min at 94°C; (2) 30 cycles of; (i) Denaturation for 5 s at 94°C; (ii) Annealing for 20 s at 59°C; (iii) Extension for 30 s at 72°C; (3) Final extension at 72°C for 5 min