An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of Pseudomonas aeruginosa isolates

Table 2 PCR parameters for the detection of genes encoding TTSS components

Gene	Forward & reverse primer sequences (all primers listed 5' to 3')	PCR conditions
exoS	S1: ATGGCGTGTTCCGAGTCA	Annealing temperature step-down 1°C/cycle (8 cycles) from 62°C to 55°C, followed by 25 cycles at 54°C. All extensions carried out at 72°C for 90 sec.
	S2: AGGTGTCGGTTCGTGACGTCT
exoT	T1: GAGAGGCTGGCGAAGGATCAC
	T2: CCAGGTCCAGAGCATCGAGCA
exoU	U1: GGCACATATCTCCGGTTCCTTC
	U2: TCAACTCAGCTGCCAACCATGC
pcrV	V1: ATGGAAGTCAGAAACCTTAATGCCG	Annealing temperature step-down 1°C/cycle (8 cycles) from 58°C to 51°C, followed by 25 cycles at 50°C. All extensions carried out at 72°C for 75 sec.
	V2: CTAGATCGCGCTGAGAATGTC
popD	D1: ATGATCGACACGCAATATTCCC
	D2: TCAGACCACTCCGGCCGCCGCA

ISSN: 1476-0711