Comparison of a commercial real-time PCR assay for tcdB detection to a cell culture cytotoxicity assay and toxigenic culture for direct detection of toxin-producing Clostridium difficile in clinical samples

Journal of Clinical Microbiology

Volumn 47, Issue 2, 2009, Pages 373-378

  Stamper, Paul D ^a   Alcabasa, Romina ^b   Aird, Deborah ^b   Babiker, Wisal ^b   Wehrlin, Jennifer ^b   Ikpeama, Ijeoma ^b   Carroll, Karen C ^a,b,c  

^a JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b JOHNS HOPKINS HOSPITAL   (United States)

^c DEPARTMENT OF PATHOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CEFOXITIN; CLOSTRIDIUM DIFFICILE TOXIN B; CYCLOSERINE; CYTOTOXIN; FRUCTOSE; BACTERIAL PROTEIN; BACTERIAL TOXIN; TOXB PROTEIN, CLOSTRIDIUM DIFFICILE;

ARTICLE; BACTERIUM CULTURE; CELL ASSAY; CELL CULTURE; CLOSTRIDIUM DIFFICILE; COMPARATIVE STUDY; CONTROLLED STUDY; FECES ANALYSIS; HUMAN; INOCULATION; MAJOR CLINICAL STUDY; NONHUMAN; PRIORITY JOURNAL; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; SENSITIVITY AND SPECIFICITY; ADULT; ANIMAL; CHEMISTRY; CULTURE MEDIUM; CULTURE TECHNIQUE; ENTEROCOLITIS; EVALUATION; FECES; GENETICS; ISOLATION AND PURIFICATION; METHODOLOGY; MICROBIOLOGY; PATHOGENICITY; POLYMERASE CHAIN REACTION; SERODIAGNOSIS;

BACTERIA (MICROORGANISMS); CLOSTRIDIUM DIFFICILE; EQUIDAE;

ADULT; ANIMALS; BACTERIAL PROTEINS; BACTERIAL TOXINS; CELL CULTURE TECHNIQUES; CLOSTRIDIUM DIFFICILE; CULTURE MEDIA; ENTEROCOLITIS; FECES; HUMANS; NEUTRALIZATION TESTS; POLYMERASE CHAIN REACTION; SENSITIVITY AND SPECIFICITY;

EID: 60849136597     PISSN: 00951137     EISSN: None     Source Type: Journal    
DOI: 10.1128/JCM.01613-08     Document Type: Article

References (32)

1. Akerlund, T., I. Persson, M. Unemo, T. Norén, B. Svenungsson, M. Wullt, and L. G. Burman. 2008. Increased sporulation rate of epidemic Clostridium difficile type 027/NAP1. J. Clin. Microbiol. 46:1530-1533.
2. Bartlett, J. G. 2006. Narrative review: the new epidemic of Clostridium difficile-associated enteric disease. Ann. Intern. Med. 145:758-764.
3. Bélanger, S. D., M. Boissinot, N. Clairoux, F. J. Picard, and M. G. Bergeron. 2003. Rapid detection of Clostridium difficile in feces by real-time PCR. J. Clin. Microbiol. 41:730-734.
4. Benson, L., X. Song, J. Campos, and N. Singh. 2007. Changing epidemiology of Clostridium difficile-associated disease in children. Infect. Control Hosp. Epidemiol. 28:1233-1235.
5. Bouza, E., T. Peláez, R. Alonso, P. Catalán, P. Muñoz, and M. R. Créixems. 2001. "Second-look" cytotoxicity: an evaluation of culture plus cytotoxin assay of Clostridium difficile isolates in the laboratory diagnosis of CDAD. J. Hosp. Infect. 48:233-237.
6. Centers for Disease Control and Prevention (CDC). 2008. Surveillance for community-associated Clostridium difficile - Connecticut, 2006. MMWR Morb. Mortal. Wkly. Rep. 57:340-343.
7. Delmée, M., J. Van Broeck, A. Simon, M. Janssens, and V. Avesani. 2005. Laboratory diagnosis of Clostridium difficile-associated diarrhoea: a plea for culture. J. Med. Microbiol. 54:187-191.
8. Drudy, D., N. Harnedy, S. Fanning, M. Hannan, and L. Kyne. 2007. Emergence and control of fluoroquinolone-resistant, toxin A-negative, toxin Bpositive Clostridium difficile. Infect. Control Hosp. Epidemiol. 28:932-940.
9. Fenner, L., A. F. Widmer, G. Goy, S. Rudin, and R. Frei. 2008. Rapid and reliable diagnostic algorithm for detection of Clostridium difficile. J. Clin. Microbiol. 46:328-330.
10. Gilligan, P. H. 2008. Is a two-step glutamate dehydrogenase antigen-cytotoxicity neutralization assay algorithm superior to the premier toxin A and B enzyme immunoassay for laboratory detection of Clostridium difficile? J. Clin. Microbiol. 46:1523-1525.
11. Guilbault, C., A. C. Labbé, L. Poirier, L. Busque, C. Béliveau, and M. Laverdière. 2002. Development and evaluation of a PCR method for detection of the Clostridium difficile toxin B gene in stool specimens. J. Clin. Microbiol. 40:2288-2290.
12. Hubert, B., V. G. Loo, A. M. Bourgault, L. Poirier, A. Dascal, E. Fortin, M. Dionne, and M. Lorange. 2007. A portrait of the geographic dissemination of the Clostridium difficile North American pulsed-field type 1 strain and the epidemiology of C. difficile-associated disease in Québec. Clin. Infect. Dis. 44:238-244.
13. Kyne, L., M. B. Hamel, R. Polavaram, and C. P. Kelly. 2002. Health care costs and mortality associated with nosocomial diarrhea due to Clostridium difficile. Clin. Infect. Dis. 34:346-353.
14. Loo, V. G., L. Poirier, M. A. Miller, M. Oughton, M. D. Libman, S. Michaud, A. M. Bourgault, T. Nguyen, C. Frenette, M. Kelly, A. Vibien, P. Brassard, S. Fenn, K. Dewar, T. J. Hudson, R. Horn, P. René, Y. Monczak, and A. Dascal. 2006. 2005. A predominantly clonal multi-institutional outbreak of Clostridium difficile-associated diarrhea with high morbidity and mortality. N. Engl. J. Med. 353:2442-2449. (Erratum, 354:2200, 2006.)
15. Lyerly, D. M., L. M. Neville, D. T. Evans, J. Fill, S. Allen, W. Greene, R. Sautter, P. Hnatuck, D. J. Torpey, and R. Schwalbe. 1998. Multicenter evaluation of the Clostridium difficile TOX A/B test. J. Clin. Microbiol. 36:184-190.
16. McDonald, L. C., G. E. Killgore, A. Thompson, R. C. Owens, Jr., S. V. Kazakova, S. P. Sambol, S. Jonson, and D. N. Gerding. 2005. An epidemic, toxin gene-variant strain of Clostridium difficile. N. Engl. J. Med. 353:2433-2441.
17. Mehlig, M., M. Moos, V. Braun, B. Kalt, D. E. Mahony, and C. von Eichel-Streiber. 2001. Variant toxin B and a functional toxin A produced by Clostridium difficile C34. FEMS Microbiol. Lett. 198:171-176.
18. Merz, C. S., C. Kramer, M. Forman, L. Gluck, K. Mills, K. Senft, I. Steiman, N. Wallace, and P. Charache. 1994. Comparison of four commercially available rapid enzyme immunoassays with cytotoxin assay for detection of Clostridium difficile toxin(s) from stool specimens. J. Clin. Microbiol. 32:1142-1147.
19. Musher, D. M., A. Manhas, P. Jain, F. Nuila, A. Waqar, N. Logan, B. Marino, and E. A. Graviss. 2007. Detection of Clostridium difficile toxin: comparison of enzyme immunoassay results with results obtained by cytotoxicity assay. J. Clin. Microbiol. 45:2737-2739.
20. O'Brien, J. A., B. J. Lahue, J. J. Caro, and D. M. Davidson. 2007. The emerging infectious challenge of Clostridium difficile-associated disease in Massachusetts hospitals: clinical and economic consequences. Infect. Control Hosp. Epidemiol. 28:1219-1227.
21. Perelle, S., M. Gibert, P. Bourlioux, G. Corthier, and M. R. Popoff. 1997. Production of a complete binary toxin (actin-specific ADP-ribosyltransferase) by Clostridium difficile CD196. Infect. Immun. 65:1402-1407.
22. Peterson, L. R., R. U. Manson, S. M. Paule, D. M. Hacek, A. Robicsek, R. B. Thomson, Jr., and K. L. Kaul. 2007. Detection of toxigenic Clostridium difficile in stool samples by real-time polymerase chain reaction for the diagnosis of C. difficile-associated diarrhea. Clin. Infect. Dis. 45:1152-1160.
23. Reller, M. E., C. A. Lema, T. M. Perl, M. Cai, T. L. Ross, K. A. Speck, and K. C. Carroll. 2007. Yield of stool culture with isolate toxin testing versus a two-step algorithm including stool toxin testing for detection of toxigenic Clostridium difficile. J. Clin. Microbiol. 45:3601-3605.
24. Rupnik, M. 2008. Heterogeneity of large clostridial toxins: importance of Clostridium difficile toxinotypes. FEMS Microbiol. Rev. 32:541-555.
25. Shanholtzer, C. J., K. E. Willard, J. J. Holter, M. M. Olson, D. N. Gerding, and L. R. Peterson. 1992. Comparison of the VIDAS Clostridium difficile toxin A immunoassay with C. difficile culture and cytotoxin and latex tests. J. Clin. Microbiol. 30:1837-1840.
26. Sloan, L. M., B. J. Duresko, D. R. Gustafson, and J. E. Rosenblatt. 2008. Comparison of real-time PCR for detection of the tcdC gene with four toxin immunoassays and culture in diagnosis of Clostridium difficile infection. J. Clin. Microbiol. 46:1996-2001.
27. Snell, H., M. Ramos, S. Longo, M. John, and Z. Hussain. 2004. Performance of the TechLab C. DIFF CHEK-60 enzyme immunoassay (EIA) in combination with the C. difficile Tox A/B II EIA kit, the Triage C. difficile panel immunoassay, and a cytotoxin assay for diagnosis of Clostridium difficile-associated diarrhea. J. Clin. Microbiol. 42:4863-4865.
28. Ticehurst, J. R., D. Z. Aird, L. M. Dam, A. P. Borek, J. T. Hargrove, and K. C. Carroll. 2006. Effective detection of toxigenic Clostridium difficile by a two-step algorithm including tests for antigen and cytotoxin. J. Clin. Microbiol. 44:1145-1149.
29. van den Berg, R. J., H. A. Ameen, T. Furusawa, E. C. Claas, E. R. van der Vorm, and E. J. Kuijper. 2005. Coexistence of multiple PCR-ribotype strains of Clostridium difficile in faecal samples limits epidemiological studies. J. Med. Microbiol. 54:173-179.
30. van den Berg, R. J., L. S. Bruijnesteijn van Coppenraet, H. J. Gerritsen, H. P. Endtz, E. R. van der Vorm, and E. J. Kuijper. 2005. Prospective multicenter evaluation of a new immunoassay and real-time PCR for rapid diagnosis of Clostridium difficile-associated diarrhea in hospitalized patients. J. Clin. Microbiol. 43:5338-5340.
31. van den Berg, R. J., E. J. Kuijper, L. E. van Coppenraet, and E. C. Claas. 2006. Rapid diagnosis of toxinogenic Clostridium difficile in faecal samples with internally controlled real-time PCR. Clin. Microbiol. Infect. 12:184-186.
32. van den Berg, R. J., N. Vaessen, H. P. Endtz, T. Schülin, E. R. van der Vorm, and E. J. Kuijper. 2007. Evaluation of real-time PCR and conventional diagnostic methods for the detection of Clostridium difficile-associated diarrhoea in a prospective multicentre study. J. Med. Microbiol. 56:36-42.