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Volume 9, Number 1—January 2003

Dispatch

Single Multiplex Polymerase Chain Reaction To Detect Diverse Loci Associated with Diarrheagenic Escherichia coli

Catalina López-Saucedo*, Jorge F. Cerna*, Nicolas Villegas-Sepulveda*, Rocío Thompson*, F. Raul Velazquez†, Javier Torres†, Phillip I. Tarr‡§, and Teresa Estrada-García*Comments to Author 
Author affiliations: *Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, México D.F., México; †Hospital de Pediatría, México D.F., México; ‡Children’s Hospital and Regional Medical Center, Seattle, Washington, USA; §University of Washington School of Medicine, Seattle, Washington, USA

Main Article

Table 1

Prototypes and reference strains of ETEC, EPEC, EIEC, and STEC tested in multiplex PCR by using specific oligonucleotide primers for several locusa

E. coli category tested
strains and serotypes Locus Primers Amplicon
size (bp) Primer (pMol)
in mix
ETEC
H10407 O78:H11b (7)
E9034A O8:H9 (8)
B2C O6:H16 (8)
E8775A O25:H42c (9)
lt
F:5´GGC GAC AGA TTA TAC CGT GC3´ (4)
R:5´ CGG TCT CTA TAT TCC CTG TT3´ (4)
440
5.0
ETEC
H10407 O78:H11b (7)
E9034A O8:H9 (8)
B2C O6:H16 (8)
E8775A O25:H42c (9)
st
F:5´ATT TTT CTT TCT GTA TTG TCT T3´ (4)
R:5´CAC CCG GTA CAA GCA GGA TT3´ (4)
191
6.47
EPEC
E2348-69 O127:H6b (10)
B171-8 O111:NM (10)
659-79 O119:H6 (10)
E851/71 O142:H6 (10)
bfpA
F:5´AAT GGT GCT TGC GCT TGC TGC3´ (5)
R:5´ GCC GCT TTA TCC AAC CTG GTA3´ (5)
324
2.5
EPEC
E2348-69 O127:H6b (10)
B171-8 O111:NM (10)
659-79 O119:H6 (10)
E851/71 O142:H6 (10)
STEC
EDL933 O157:H7b (11)
TB334C O85:NM (12)
TB285A O126:H2 (12)
TB226A O11:HN(12)
eaeA
F:5´ GAC CCG GCA CAA GCA TAA GC3´ (6)
R:5´CCA CCT GCA GCA ACA AGA GG3´ (6)
384
3.88
STEC
EDL933 O157:H7b (11)
TB334C O85:NM (12)
TB285A O126:H2 (12)
TB226A O11:HN (12)
stx1
F:5´CTG GAT TTA ATG TCG CAT AGT G3´ d (GenBank accession no. M17358)
R:5´AGA ACG CCC ACT GAG ATC ATC3´ (6)
150
3.88
STEC
EDL933 O157:H7b (11)
TB226A O11:HN (12)
stx2
F:5´GGC ACT GTC TGA AAC TGC TCC3´ (6)
R:5´TCG CCA GTT ATC TGA CAT TCT G3´ (6)
255
2.5
EIEC
E11 O124NMb (13)
O124:H30 (14)
O136:NM (14)
O143:NM (14) ial F:5´GGT ATG ATG ATG ATG AGT CCA 3´ d
(GenBank accession no. D13663)
R:5´ GGA GGC CAA CAA TTA TTT CC 3´d 650 10.25

aE. coli, Escherichia coli; ETEC, enterotoxigenic E. coli; EPEC, enteropathogenic E. coli; EIEC,
enteroinvasive E. coli; STEC, Shiga-toxin–producing E. coli; PCR, polymerase chain reaction.
bE. coli prototype strains.
cDonated by the Public Health Laboratory Service, Central Health Laboratory, London, United
Kingdom.
dThese primers were designed by us from the gene bank sequences.

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