• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Mar 1995; 33(3): 519–524.
PMCID: PMC227983

Direct detection of verotoxin-producing Escherichia coli in stool samples by PCR.


A method for the rapid detection of verotoxin-producing Escherichia coli in stool samples by PCR was evaluated. Verotoxin-1 and verotoxin-2 genes in DNA extracted directly from stool samples were amplified with oligonucleotide primers. Stools spiked with control organisms, E. coli C600 (H19B) (verotoxin-1) or E. coli C600 (933W) (verotoxin-2), demonstrated that verotoxin-1-containing organisms could be detected at 10(2) CFU per 0.1 g of stool and verotoxin-2-containing organisms could be detected at 10(7) CFU per 0.1 g of stool. Testing of stool samples from patients with diarrhea showed a high concordance between PCR positivity and the presence of verotoxin-producing E. coli, determined by isolation of serotype O157:H7 on sorbitol-MacConkey medium (34 of 35 stool samples) or by colony blots with gene probes (19 of 21 stool samples). Conversely, only 1 of 20 (5.0%) stool samples that were O157:H7 culture negative and colony blot negative and that contained free verotoxin only was positive by PCR. As well, only 4 of 145 (2.8%) stool samples that were negative for serotype O157:H7 or free verotoxin were PCR positive. PCR of DNA extracted directly from stool samples provides a rapid method for the detection of stool samples containing verotoxin-producing E. coli compared with colony blot testing.

Full Text

The Full Text of this article is available as a PDF (272K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allard A, Girones R, Juto P, Wadell G. Polymerase chain reaction for detection of adenoviruses in stool samples. J Clin Microbiol. 1990 Dec;28(12):2659–2667. [PMC free article] [PubMed]
  • Basta M, Karmali M, Lingwood C. Sensitive receptor-specified enzyme-linked immunosorbent assay for Escherichia coli verocytotoxin. J Clin Microbiol. 1989 Jul;27(7):1617–1622. [PMC free article] [PubMed]
  • Begum D, Strockbine NA, Sowers EG, Jackson MP. Evaluation of a technique for identification of Shiga-like toxin-producing Escherichia coli by using polymerase chain reaction and digoxigenin-labeled probes. J Clin Microbiol. 1993 Dec;31(12):3153–3156. [PMC free article] [PubMed]
  • Brian MJ, Frosolono M, Murray BE, Miranda A, Lopez EL, Gomez HF, Cleary TG. Polymerase chain reaction for diagnosis of enterohemorrhagic Escherichia coli infection and hemolytic-uremic syndrome. J Clin Microbiol. 1992 Jul;30(7):1801–1806. [PMC free article] [PubMed]
  • Brown JE, Echeverria P, Taylor DN, Seriwatana J, Vanapruks V, Lexomboon U, Neill RN, Newland JW. Determination by DNA hybridization of Shiga-like-toxin-producing Escherichia coli in children with diarrhea in Thailand. J Clin Microbiol. 1989 Feb;27(2):291–294. [PMC free article] [PubMed]
  • Calderwood SB, Mekalanos JJ. Iron regulation of Shiga-like toxin expression in Escherichia coli is mediated by the fur locus. J Bacteriol. 1987 Oct;169(10):4759–4764. [PMC free article] [PubMed]
  • Downes FP, Barrett TJ, Green JH, Aloisio CH, Spika JS, Strockbine NA, Wachsmuth IK. Affinity purification and characterization of Shiga-like toxin II and production of toxin-specific monoclonal antibodies. Infect Immun. 1988 Aug;56(8):1926–1933. [PMC free article] [PubMed]
  • Downes FP, Green JH, Greene K, Strockbine N, Wells JG, Wachsmuth IK. Development and evaluation of enzyme-linked immunosorbent assays for detection of shiga-like toxin I and shiga-like toxin II. J Clin Microbiol. 1989 Jun;27(6):1292–1297. [PMC free article] [PubMed]
  • Frankel G, Riley L, Giron JA, Valmassoi J, Friedmann A, Strockbine N, Falkow S, Schoolnik GK. Detection of Shigella in feces using DNA amplification. J Infect Dis. 1990 Jun;161(6):1252–1256. [PubMed]
  • Gannon VP, King RK, Kim JY, Thomas EJ. Rapid and sensitive method for detection of Shiga-like toxin-producing Escherichia coli in ground beef using the polymerase chain reaction. Appl Environ Microbiol. 1992 Dec;58(12):3809–3815. [PMC free article] [PubMed]
  • Gouvea V, Allen JR, Glass RI, Fang ZY, Bremont M, Cohen J, McCrae MA, Saif LJ, Sinarachatanant P, Caul EO. Detection of group B and C rotaviruses by polymerase chain reaction. J Clin Microbiol. 1991 Mar;29(3):519–523. [PMC free article] [PubMed]
  • Gouvea V, Glass RI, Woods P, Taniguchi K, Clark HF, Forrester B, Fang ZY. Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens. J Clin Microbiol. 1990 Feb;28(2):276–282. [PMC free article] [PubMed]
  • Griffin PM, Tauxe RV. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol Rev. 1991;13:60–98. [PubMed]
  • Hii JH, Gyles C, Morooka T, Karmali MA, Clarke R, De Grandis S, Brunton JL. Development of verotoxin 2- and verotoxin 2 variant (VT2v)-specific oligonucleotide probes on the basis of the nucleotide sequence of the B cistron of VT2v from Escherichia coli E32511 and B2F1. J Clin Microbiol. 1991 Dec;29(12):2704–2709. [PMC free article] [PubMed]
  • Hornes E, Wasteson Y, Olsvik O. Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction. J Clin Microbiol. 1991 Nov;29(11):2375–2379. [PMC free article] [PubMed]
  • Jackson MP. Detection of Shiga toxin-producing Shigella dysenteriae type 1 and Escherichia coli by using polymerase chain reaction with incorporation of digoxigenin-11-dUTP. J Clin Microbiol. 1991 Sep;29(9):1910–1914. [PMC free article] [PubMed]
  • Jiang X, Wang J, Graham DY, Estes MK. Detection of Norwalk virus in stool by polymerase chain reaction. J Clin Microbiol. 1992 Oct;30(10):2529–2534. [PMC free article] [PubMed]
  • Johnson WM, Pollard DR, Lior H, Tyler SD, Rozee KR. Differentiation of genes coding for Escherichia coli verotoxin 2 and the verotoxin associated with porcine edema disease (VTe) by the polymerase chain reaction. J Clin Microbiol. 1990 Oct;28(10):2351–2353. [PMC free article] [PubMed]
  • Karch H, Meyer T. Single primer pair for amplifying segments of distinct Shiga-like-toxin genes by polymerase chain reaction. J Clin Microbiol. 1989 Dec;27(12):2751–2757. [PMC free article] [PubMed]
  • Karch H, Meyer T, Rüssmann H, Heesemann J. Frequent loss of Shiga-like toxin genes in clinical isolates of Escherichia coli upon subcultivation. Infect Immun. 1992 Aug;60(8):3464–3467. [PMC free article] [PubMed]
  • Karmali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989 Jan;2(1):15–38. [PMC free article] [PubMed]
  • Karmali MA, Petric M, Lim C, Fleming PC, Arbus GS, Lior H. The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli. J Infect Dis. 1985 May;151(5):775–782. [PubMed]
  • Kleanthous H, Fry NK, Smith HR, Gross RJ, Rowe B. The use of sorbitol-MacConkey agar in conjunction with a specific antiserum for the detection of Vero cytotoxin-producing strains of Escherichia coli O 157. Epidemiol Infect. 1988 Oct;101(2):327–335. [PMC free article] [PubMed]
  • MacLeod DL, Gyles CL. Effects of culture conditions on yield of Shiga-like toxin-IIv from Escherichia coli. Can J Microbiol. 1989 Jun;35(6):623–629. [PubMed]
  • Newland JW, Neill RJ. DNA probes for Shiga-like toxins I and II and for toxin-converting bacteriophages. J Clin Microbiol. 1988 Jul;26(7):1292–1297. [PMC free article] [PubMed]
  • Olive DM. Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase. J Clin Microbiol. 1989 Feb;27(2):261–265. [PMC free article] [PubMed]
  • Paton AW, Paton JC, Goldwater PN, Manning PA. Direct detection of Escherichia coli Shiga-like toxin genes in primary fecal cultures by polymerase chain reaction. J Clin Microbiol. 1993 Nov;31(11):3063–3067. [PMC free article] [PubMed]
  • Pollard DR, Johnson WM, Lior H, Tyler SD, Rozee KR. Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction. J Clin Microbiol. 1990 Mar;28(3):540–545. [PMC free article] [PubMed]
  • Pollard DR, Johnson WM, Lior H, Tyler SD, Rozee KR. Differentiation of Shiga toxin and Vero cytotoxin type 1 genes by polymerase chain reaction. J Infect Dis. 1990 Nov;162(5):1195–1198. [PubMed]
  • Ramotar K, Boyd B, Tyrrell G, Gariepy J, Lingwood C, Brunton J. Characterization of Shiga-like toxin I B subunit purified from overproducing clones of the SLT-I B cistron. Biochem J. 1990 Dec 15;272(3):805–811. [PMC free article] [PubMed]
  • Saulnier P, Andremont A. Detection of genes in feces by booster polymerase chain reaction. J Clin Microbiol. 1992 Aug;30(8):2080–2083. [PMC free article] [PubMed]
  • Seriwatana J, Brown JE, Echeverria P, Taylor DN, Suthienkul O, Newland J. DNA probes to identify Shiga-like toxin I- and II-producing enteric bacterial pathogens isolated from patients with diarrhea in Thailand. J Clin Microbiol. 1988 Aug;26(8):1614–1615. [PMC free article] [PubMed]
  • Smith HR, Scotland SM. Vero cytotoxin-producing strains of Escherichia coli. J Med Microbiol. 1988 Jun;26(2):77–85. [PubMed]
  • Strockbine NA, Marques LR, Holmes RK, O'Brien AD. Characterization of monoclonal antibodies against Shiga-like toxin from Escherichia coli. Infect Immun. 1985 Dec;50(3):695–700. [PMC free article] [PubMed]
  • Tyler SD, Johnson WM, Lior H, Wang G, Rozee KR. Identification of verotoxin type 2 variant B subunit genes in Escherichia coli by the polymerase chain reaction and restriction fragment length polymorphism analysis. J Clin Microbiol. 1991 Jul;29(7):1339–1343. [PMC free article] [PubMed]
  • Widjojoatmodjo MN, Fluit AC, Torensma R, Verdonk GP, Verhoef J. The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol. 1992 Dec;30(12):3195–3199. [PMC free article] [PubMed]
  • Wilde J, Eiden J, Yolken R. Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions. J Clin Microbiol. 1990 Jun;28(6):1300–1307. [PMC free article] [PubMed]
  • Willshaw GA, Scotland SM, Smith HR, Rowe B. Properties of Vero cytotoxin-producing Escherichia coli of human origin of O serogroups other than O157. J Infect Dis. 1992 Oct;166(4):797–802. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...