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Volume 10, Number 10—October 2004
Research

Escherichia coli and Community-acquired Gastroenteritis, Melbourne, Australia

Roy M. Robins-Browne*†Comments to Author , Anne-Marie Bordun*†, Marija Tauschek*, Vicki R. Bennett-Wood*†, Jacinta Russell†, Frances Oppedisano†, Nicole A. Lister*, Karl A. Bettelheim*, Christopher K. Fairley*, Martha I. Sinclair‡, and Margaret E. Hellard‡1
Author affiliations: *University of Melbourne, Melbourne, Victoria, Australia; †Murdoch Childrens Research Institute, Melbourne, Victoria, Australia; ‡Monash University, Melbourne, Victoria, Australia

Main Article

Table 1

Characteristics of the polymerase chain reaction (PCR) primers used to detect pathogenic Escherichia coli in mixed culture

Target gene or virulence factor Primera Primer sequence (5′ to 3′) PCR program (30 cycles)b Product size (bp) Reference
eae 11
21 GACCCGGCACAAGCATAAGC
CCACCTGCAGCAACAAGAGG 95°C, 30 s; 54°C, 90 s; 72°C, 90 s 384 (8)
ehxA 11
21 GCATCATCAAGCGTACGTTCC
AATGAGCCAAGCTGGTTAAGCT 534 (8)
stx1 12
22 ATAAATCGCCATTCGTTGACTAC
AGAACGCCCACTGAGATCATC 95°C, 30 s; 52°C, 60 s; 72°C, 60 sc 180 (8)
stx2 12
22 GGCACTGTCTGAAACTGCTCC
TCGCCAGTTATCTGACATTCTG 255 (8)
ltA 13
23 GGCGACAGATTATACCGTGC
CCGAATTCTGTTATATATGTC 94°C, 60 s; 50°C, 60 s; 72°C, 120 sc 696 (9)
st1A 13
23 TCTGTATTATCTTTCCCCTC
ATAACATCCAGCACAGGC 186 (9)
ipaC 1
2 CAGCAGATTGCAGCGCATAT
CAAGAGCAGATGCATAACGC 94°C, 30 s; 59°C, 90 s; 72°C, 90 s 811 This study
bfpA 1
2 ATTGAATCTGCAATGGTGC
ATAGCAGTCGATTTAGCAGCC 95°C, 40 s; 55°C, 40 s; 72°C, 40 s 461 This study
pCVD432 1
2 CTGGCGAAAGACTGTATCAT
CAATGTATAGAAATCCGCTGTT 94°C, 40 s; 53°C, 60 s; 72°C, 60 s 630 (10)
aggA 1
2 ATGCATTACTTTGGGTTTAG
TCAACCTTGACACTTGCC 94°C, 60 s; 50°C, 60 s; 72°C, 120 sc 414 This study
lacZ 1
2 TGATTGAAGCAGAAGCCTGC
CGCCAATCCACATCTGTGAA 94°C, 30 s; 59°C, 90 s; 72°C, 90 s 1,350 This study

aPrimers with the same superscript were used in duplex PCRs.
bBefore the first cycle, the sample was denatured for 10 min at 95°C; after the last cycle, the sample was extended for 8 min at 72°C.
c35 cycles.

Main Article

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1Current affiliation: Burnet Institute, Melbourne, Victoria, Australia.

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Page updated: April 11, 2011
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