1887

Abstract

Summary

A rapid assay was developed for detection of the enterotoxin gene in stool specimens by means of the polymerase chain reaction (PCR). The PCR primers amplified a 63-bp repetitive sequence of the enterotoxin gene, thereby generating a distinctive ladder pattern of DNA bands following electrophoresis. Crude DNA extracts from stools containing produced one (63-bp) or more bands of the characteristic ladder. Of 172 stool specimens from 58 patients, 37 gave positive results by culture (15 specimens) or cytotoxin assay (36 specimens). When 36 available “positive” specimens were tested by the PCR assay, 34 (94%) gave positive results-24 by direct testing, and 10 after extraction of DNA by the Qiagen procedure. Insufficient material of the remaining two specimens was available for DNA extraction. Of 21 stools “negative” for by culture or cytotoxin assay, one gave a positive result by PCR and seven produced atypical bands. The rapid PCR detection technique for was more sensitive than standard culture, and of a sensitivity similar to cytotoxin testing. The method has the potential for adoption in routine laboratory practice.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-38-5-384
1993-05-01
2024-05-03
Loading full text...

Full text loading...

/deliver/fulltext/jmm/38/5/medmicro-38-5-384.html?itemId=/content/journal/jmm/10.1099/00222615-38-5-384&mimeType=html&fmt=ahah

References

  1. Bartlett JG, Chang TW, Gurwith M, Gorbach SL, Onderdonk AB. Antibiotic-associated pseudomembranous colitis due to toxin-producing clostridia. N Engl J Med 1978; 298:531–534
    [Google Scholar]
  2. George RH, Symonds JM, Dimock F et al. Identification of Clostridium difficile as a cause of pseudomembranous colitis. BMJ 1978; 1:695
    [Google Scholar]
  3. Banno Y, Kobayashi T, Watanabe K, Ueno K, Nozawa Y. Two toxins (D-l and D-2) of Clostridium difficile causing antibiotic-associated colitis: purification and some characterization. Biochem Int 1981; 2:629–635
    [Google Scholar]
  4. Sullivan NM, Pellett S, Wilkins TD. Purification and char acterization of toxins A and B of Clostridium difficile . Infect Immun 1982; 35:1032–1040
    [Google Scholar]
  5. Wilkins TD. Role of Clostridium difficile toxins in disease. Gastroenterology 1987; 93:389–391
    [Google Scholar]
  6. Walker RC, Ruane PJ, Rosenblatt JE et al. Comparison of culture, cytotoxicity assays and enzyme-linked sorbent assay for toxin A and toxin B in the diagnosis of Clostridium difficile-related enteric disease. Diagn Microbiol Infect Dis 1986; 5:61–69
    [Google Scholar]
  7. Wren BW, Clayton CL, Tabaqchali S. Nucleotide sequence of Clostridium difficile toxin A gene fragment and detection of toxigenic strains by polymerase chain reaction. FEMS Microbiol Lett 1990; 58:1–6
    [Google Scholar]
  8. Dove CH, Wang S-Z, Price SB et al. Molecular characterization of the Clostridium difficile toxin A gene. Infect Immun 1990; 58:480–488
    [Google Scholar]
  9. Cato EP, George WL, Finegold SM. Genus Clostridium. In Sneath PHA, Mair NS, Sharpe ME, Holt JG. eds Bergey’s manual of systematic bacteriology, vol 2. Baltimore: Williams and Wilkins; 1986:1141–1200
    [Google Scholar]
  10. Chang TW, Lauerman M, Bartlett SG. Cytotoxicity assay in antibiotic-associated colitis. J Infect Dis 1979; 140:765–770
    [Google Scholar]
  11. Khan G, Kangro HO, Coates PJ, Heath JB. Inhibitory effects of urine on the polymerase chain reaction for cytomegalovirus DNA. J Clin Pathol 1991; 44:360–365
    [Google Scholar]
  12. Corthier G, Muller MC. Emergence in gnotobiotic mice of non-toxigenic clones of Clostridium difficile from a toxigenic one. Infect Immun 1988; 56:1500–1504
    [Google Scholar]
  13. Wren BW, Clayton CL, Tabaqchali S. Rapid identification of toxigenic Clostridium difficile by polymerase chain reaction. Lancet 1990; 335:423
    [Google Scholar]
  14. Wren BW, Clayton CL, Castledine NB, Tabaqchali S. Identification of toxigenic Clostridium difficile strains by using a toxin A gene-specific probe. J Clin Microbiol 1990; 28:1808–1812
    [Google Scholar]
  15. Kato N, Ou C-Y, Kato H et al. Identification of toxigenic Clostridium difficile by the polymerase chain reaction. J Clin Microbiol 1991; 29:33–37
    [Google Scholar]
  16. Gumerlock PH, Tang YJ, Meyers FJ, Silva J. Use of the polymerase chain reaction for the specific and direct detection of Clostridium difficile in human faeces. Rev Infect Dis 1991; 13:1053–1060
    [Google Scholar]
  17. Wren BW, Pallen MJ, Tabaqchali S. Beneficial effects of #x201C;jumping PCR” in diagnostic procedures. Lancet 1990; 1:1540
    [Google Scholar]
  18. Olive DM. Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase. J Clin Microbiol 1989; 27:261–265
    [Google Scholar]
  19. Gouvea V, Glass RI, Woods P et al. Polymerase chain reaction amplification and typing of rotavirus nucleic acid from stool specimens. J Clin Microbiol 1990; 28:276–282
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-38-5-384
Loading
/content/journal/jmm/10.1099/00222615-38-5-384
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error