Development and validation of digital enzyme-linked immunosorbent assays for ultrasensitive detection and quantification of clostridium difficile toxins in stool

Journal of Clinical Microbiology

Volumn 53, Issue 10, 2015, Pages 3204-3212

  Song, Linan ^a   Zhao, Mingwei ^a   Duffy, David C ^a   Hansen, Joshua ^b   Shields, Kelsey ^b   Wungjiranirun, Manida ^b   Chen, Xinhua ^b   Xu, Hua ^b   Leffler, Daniel A ^b   Sambol, Susan P ^c,d   Gerding, Dale N ^c,d   Kelly, Ciarán P ^b   Pollock, Nira R ^b,e  

^a QUANTERIX CORPORATION   (United States)

^b BETH ISRAEL DEACONESS MEDICAL CENTER   (United States)

^c VETERANS AFFAIRS MEDICAL CENTER   (United States)

^d LOYOLA UNIVERSITY CHICAGO   (United States)

^e BOSTON CHILDREN S HOSPITAL   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CLOSTRIDIUM DIFFICILE TOXIN A; CLOSTRIDIUM DIFFICILE TOXIN B; BACTERIAL PROTEIN; BACTERIAL TOXIN; ENTEROTOXIN; TCDA PROTEIN, CLOSTRIDIUM DIFFICILE; TOXB PROTEIN, CLOSTRIDIUM DIFFICILE;

ADULT; ARTICLE; BACTERIAL STRAIN; BACTERIUM ISOLATE; CALIBRATION; CLOSTRIDIUM DIFFICILE INFECTION; CONCENTRATION (PARAMETERS); CONTROLLED STUDY; CULTURE TECHNIQUE; CYTOTOXICITY TEST; DISEASE ASSOCIATION; DISEASE SEVERITY; ENZYME LINKED IMMUNOSORBENT ASSAY; EXPERIMENTAL DESIGN; FECES ANALYSIS; HUMAN; LIMIT OF DETECTION; MAJOR CLINICAL STUDY; NONHUMAN; NUCLEIC ACID AMPLIFICATION; PEPTOCLOSTRIDIUM DIFFICILE; PRIORITY JOURNAL; PROCESS OPTIMIZATION; QUANTITATIVE ANALYSIS; REFERENCE VALUE; SENSITIVITY AND SPECIFICITY; TOXIGENIC CULTURE; TOXIN ANALYSIS; ULTRASENSITIVE DIGITAL ENZYME LINKED IMMUNOSORBENT ASSAY; VALIDATION STUDY; AGED; CHEMISTRY; CLOSTRIDIUM INFECTIONS; DIARRHEA; FECES; FEMALE; LABORATORY TEST; MALE; MIDDLE AGED; PROCEDURES; VERY ELDERLY; YOUNG ADULT;

ADULT; AGED; AGED, 80 AND OVER; BACTERIAL PROTEINS; BACTERIAL TOXINS; CLINICAL LABORATORY TECHNIQUES; CLOSTRIDIUM INFECTIONS; DIARRHEA; ENTEROTOXINS; ENZYME-LINKED IMMUNOSORBENT ASSAY; FECES; FEMALE; HUMANS; MALE; MIDDLE AGED; SENSITIVITY AND SPECIFICITY; YOUNG ADULT;

EID: 84941979099     PISSN: 00951137     EISSN: 1098660X     Source Type: Journal    
DOI: 10.1128/JCM.01334-15     Document Type: Article

References (49)

1. Freeman J, Bauer MP, Baines SD, Corver J, Fawley WN, Goorhuis B, Kuijper EJ, Wilcox MH. 2010. The changing epidemiology of Clostridium difficile infections. Clin Microbiol Rev 23:529-549. http://dx. doi. org/10 . 1128/CMR. 00082-09.
2. Cohen SH, Gerding DN, Johnson S, Kelly CP, Loo VG, McDonald LC, Pepin J, Wilcox MH, Society for Healthcare Epidemiology of America, Infectious Diseases Society of America. 2010. Clinical practice guidelines for Clostridium difficile infection in adults: 2010 update by the Society for Healthcare Epidemiology of America (SHEA) and the Infectious Diseases Society of America (IDSA). Infect Control Hosp Epidemiol 31:431-455. http://dx. doi. org/10. 1086/651706.
3. Rupnik M, Wilcox MH, Gerding DN. 2009. Clostridium difficile infection: new developments in epidemiology and pathogenesis. Nat Rev Microbiol 7:526-536. http://dx. doi. org/10. 1038/nrmicro2164.
4. Schutze GE, Willoughby RE, Committee on Infectious Diseases. 2013. Clostridium difficile infection in infants and children. Pediatrics 131:196-200. http://dx. doi. org/10. 1542/peds. 2012-2992.
5. Magill SS, Edwards JR, Bamberg W, Beldavs ZG, Dumyati G, Kainer MA, Lynfield R, Maloney M, McAllister-Hollod L, Nadle J, Ray SM, Thompson DL, Wilson LE, Fridkin SK, Emerging Infections Program Healthcare-Associated Infections and Antimicrobial Use Prevalence Survey Team. 2014. Multistate point-prevalence survey of health careassociated infections. N Engl J Med 370:1198-1208. http://dx. doi. org/10 . 1056/NEJMoa1306801.
6. Kamboj M, Khosa P, Kaltsas A, Babady NE, Son C, Sepkowitz KA. 2011. Relapse versus reinfection: surveillance of Clostridium difficile infection. Clin Infect Dis 53:1003-1006. http://dx. doi. org/10. 1093/cid/cir643.
7. McDonald LC, Killgore GE, Thompson A, Owens RC, Jr, Kazakova SV, Sambol SP, Johnson S, Gerding DN. 2005. An epidemic, toxin genevariant strain of Clostridium difficile. N Engl J Med 353:2433-2441. http: //dx. doi. org/10. 1056/NEJMoa051590.
8. Loo VG, Poirier L, Miller MA, Oughton M, Libman MD, Michaud S, Bourgault AM, Nguyen T, Frenette C, Kelly M, Vibien A, Brassard P, Fenn S, Dewar K, Hudson TJ, Horn R, Rene P, Monczak Y, Dascal A. 2005. A predominantly clonal multi-institutional outbreak of Clostridium difficile-associated diarrhea with high morbidity and mortality. N Engl J Med 353:2442-2449. http://dx. doi. org/10. 1056/NEJMoa051639.
9. Lyras D, O'Connor JR, Howarth PM, Sambol SP, Carter GP, Phumoonna T, Poon R, Adams V, Vedantam G, Johnson S, Gerding DN, Rood JI. 2009. Toxin B is essential for virulence of Clostridium difficile. Nature 458:1176-1179. http://dx. doi. org/10. 1038/nature07822.
10. Kuehne SA, Cartman ST, Heap JT, Kelly ML, Cockayne A, Minton NP. 2010. The role of toxin A and toxin B in Clostridium difficile infection. Nature 467:711-713. http://dx. doi. org/10. 1038/nature09397.
11. Dubberke ER, Han Z, Bobo L, Hink T, Lawrence B, Copper S, Hoppe-Bauer J, Burnham CA, Dunne WM, Jr. 2011. Impact of clinical symptoms on interpretation of diagnostic assays for Clostridium difficile infections. J Clin Microbiol 49:2887-2893. http://dx. doi. org/10. 1128/JCM . 00891-11.
12. Planche TD, Davies KA, Coen PG, Finney JM, Monahan IM, Morris KA, O'Connor L, Oakley SJ, Pope CF, Wren MW, Shetty NP, Crook DW, Wilcox MH. 2013. Differences in outcome according to Clostridium difficile testing method: a prospective multicentre diagnostic validation study of C. difficile infection. Lancet Infect Dis 13:936-945. http://dx. doi . org/10. 1016/S1473-3099(13)70200-7.
13. He X, Wang J, Steele J, Sun X, Nie W, Tzipori S, Feng H. 2009. An ultrasensitive rapid immunocytotoxicity assay for detecting Clostridium difficile toxins. J Microbiol Methods 78:97-100. http://dx. doi. org/10. 1016 /j. mimet. 2009. 04. 007.
14. Wilcox MH, Planche T, Fang FC, Gilligan P. 2010. What is the current role of algorithmic approaches for diagnosis of Clostridium difficile infection? J Clin Microbiol 48:4347-4353. http://dx. doi. org/10. 1128/JCM . 02028-10.
15. Ota KV, McGowan KL. 2012. Clostridium difficile testing algorithms using glutamate dehydrogenase antigen and C. difficile toxin enzyme immunoassays with C. difficile Nucleic acid amplification testing increase diagnostic yield in a tertiary pediatric population. J Clin Microbiol 50: 1185-1188. http://dx. doi. org/10. 1128/JCM. 05620-11.
16. Koo HL, Van JN, Zhao M, Ye X, Revell PA, Jiang ZD, Grimes CZ, Koo DC, Lasco T, Kozinetz CA, Garey KW, DuPont HL. 2014. Real-time polymerase chain reaction detection of asymptomatic Clostridium difficile colonization and rising C. difficile-associated disease rates. Infect Control Hosp Epidemiol 35:667-673. http://dx. doi. org/10. 1086 /676433.
17. Leibowitz J, Soma VL, Rosen L, Ginocchio CC, Rubin LG. 2015. Similar proportions of stool specimens from hospitalized children with and without diarrhea test positive for Clostridium difficile. Pediatr Infect Dis J 34: 261-266. http://dx. doi. org/10. 1097/INF. 0000000000000556.
18. Kelly CP, LaMont JT. 2008. Clostridium difficile-more difficult than ever. N Engl J Med 359:1932-1940. http://dx. doi. org/10. 1056/NEJMra0707500.
19. Kelly CP, Pothoulakis C, Vavva F, Castagliuolo I, Bostwick EF, O'Keane JC, Keates S, LaMont JT. 1996. Anti-Clostridium difficile bovine immunoglobulin concentrate inhibits cytotoxicity and enterotoxicity of C. difficile toxins. Antimicrob Agents Chemother 40:373-379.
20. Meridian Bioscience, Inc. 2009. Premier toxins A & B package insert. Meridian Bioscience, Inc. , Nice, France. http://www. meridianbioscience. com/Content/Assets/Files/2. 1%20%20C. %20difficile%20Products/Pack age-Insert-Premier-Toxins-A-and-B. pdf.
21. TechLab. 2008. C. difficile Tox A/B II package insert. TechLab, Inc. , Blacksburg, VA. http://www. techlab. com/wp-content/uploads/2013/06 /t5015insert-rev-0308. pdf.
22. Crobach MJ, Dekkers OM, Wilcox MH, Kuijper EJ. 2009. European Society of Clinical Microbiology and Infectious Diseases (ESCMID): data review and recommendations for diagnosing Clostridium difficileinfection (CDI). Clin Microbiol Infect 15:1053-1066. http://dx. doi. org/10 . 1111/j. 1469-0691. 2009. 03098. x.
23. Eastwood K, Else P, Charlett A, Wilcox M. 2009. Comparison of nine commercially available Clostridium difficile toxin detection assays, a realtime PCR assay for C. difficile tcdB, and a glutamate dehydrogenase detection assay to cytotoxin testing and cytotoxigenic culture methods. J Clin Microbiol 47:3211-3217. http://dx. doi. org/10. 1128/JCM. 01082-09.
24. Akerlund T, Svenungsson B, Lagergren A, Burman LG. 2006. Correlation of disease severity with fecal toxin levels in patients with Clostridium difficile-associated diarrhea and distribution of PCR ribotypes and toxin yields in vitro of corresponding isolates. J Clin Microbiol 44:353-358. http: //dx. doi. org/10. 1128/JCM. 44. 2. 353-358. 2006.
25. Ryder AB, Huang Y, Li H, Zheng M, Wang X, Stratton CW, Xu X, Tang YW. 2010. Assessment of Clostridium difficile infections by quantitative detection of tcdB toxin by use of a real-time cell analysis system. J Clin Microbiol 48:4129-4134. http://dx. doi. org/10. 1128/JCM. 01104-10.
26. Rissin DM, Kan CW, Campbell TG, Howes SC, Fournier DR, Song L, Piech T, Patel PP, Chang L, Rivnak AJ, Ferrell EP, Randall JD, Provuncher GK, Walt DR, Duffy DC. 2010. Single-molecule enzymelinked immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nat Biotechnol 28:595-599. http://dx. doi. org/10. 1038 /nbt. 1641.
27. Rissin DM, Fournier DR, Piech T, Kan CW, Campbell TG, Song L, Chang L, Rivnak AJ, Patel PP, Provuncher GK, Ferrell EP, Howes SC, Pink BA, Minnehan KA, Wilson DH, Duffy DC. 2011. Simultaneous detection of single molecules and singulated ensembles of molecules enables immunoassays with broad dynamic range. Anal Chem 83:2279-2285. http://dx. doi. org/10. 1021/ac103161b.
28. Wilson DH, Hanlon DW, Provuncher GK, Chang L, Song L, Patel PP, Ferrell EP, Lepor H, Partin AW, Chan DW, Sokoll LJ, Cheli CD, Thiel RP, Fournier DR, Duffy DC. 2011. Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology. Clin Chem 57:1712-1721. http://dx. doi. org/10. 1373 /clinchem. 2011. 169540.
29. Song L, Hanlon DW, Chang L, Provuncher GK, Kan CW, Campbell TG, Fournier DR, Ferrell EP, Rivnak AJ, Pink BA, Minnehan KA, Patel PP, Wilson DH, Till MA, Faubion WA, Duffy DC. 2011. Single molecule measurements of tumor necrosis factor alpha and interleukin-6 in the plasma of patients with Crohn's disease. J Immunol Methods 372:177-186. http://dx. doi. org/10. 1016/j. jim. 2011. 07. 015.
30. Cepheid, Inc. 2012. GeneXpert C. difficile package insert. Cepheid, Inc.
31. Meridian Bioscience, Inc. 2011. illumigene C. difficile package insert. Meridian Bioscience, Inc. , Nice, France. http://www. meridianbioscience. com/Content/Assets/PackInsert/8-5-x-11-CLEAN-SN11176-280050-illumigene-C-difficile-REV-08-13-pdf.
32. Deshpande A, Pasupuleti V, Rolston DD, Jain A, Deshpande N, Pant C, Hernandez AV. 2011. Diagnostic accuracy of real-time polymerase chain reaction in detection of Clostridium difficile in the stool samples of patients with suspected Clostridium difficile Infection: a meta-analysis. Clin Infect Dis 53:e81-90. http://dx. doi. org/10. 1093/cid/cir505.
33. Kyne L, Warny M, Qamar A, Kelly CP. 2000. Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A. N Engl J Med 342:390-397.
34. Kan CW, Rivnak AJ, Campbell TG, Piech T, Rissin DM, Mösl M, Peterça A, Niederberger HP, Minnehan KA, Patel PP, Ferrell EP, Meyer RE, Chang L, Wilson DH, Fournier DR, Duffy DC. 2012. Isolation and detection of single molecules on paramagnetic beads using sequential fluid flows in microfabricated polymer array assemblies. Lab Chip 12:977-985. http://dx. doi. org/10. 1039/C2LC20744C.
35. Tenover FC, Akerlund T, Gerding DN, Goering RV, Bostrom T, Jonsson AM, Wong E, Wortman AT, Persing DH. 2011. Comparison of strain typing results for Clostridium difficile isolates from North America. J Clin Microbiol 49:1831-1837. http://dx. doi. org/10. 1128/JCM . 02446-10.
36. Wilson KH, Kennedy MJ, Fekety FR. 1982. Use of sodium taurocholate to enhance spore recovery on a medium selective for Clostridium difficile. J Clin Microbiol 15:443-446.
37. Clabots CR, Gerding SJ, Olson MM, Peterson LR, Gerding DN. 1989. Detection of asymptomatic Clostridium difficile carriage by an alcohol shock procedure. J Clin Microbiol 27:2386-2387.
38. Clabots CR, Johnson S, Bettin KM, Mathie PA, Mulligan ME, Schaberg DR, Peterson LR, Gerding DN. 1993. Development of a rapid and efficient restriction endonuclease analysis typing system for Clostridium difficile and correlation with other typing systems. J Clin Microbiol 31:1870-1875.
39. Petrella LA, Sambol SP, Cheknis A, Nagaro K, Kean Y, Sears PS, Babakhani F, Johnson S, Gerding DN. 2012. Decreased cure and increased recurrence rates for Clostridium difficile infection caused by the epidemic C. difficile BI strain. Clin Infect Dis 55:351-357. http://dx. doi . org/10. 1093/cid/cis430.
40. bioMérieux. 2011. Vidas C. difficile Toxin A & B package insert, bioMérieux, Marcy l'Etoile, France.
41. Meridian Bioscience, Inc. 2009. ImmunoCard toxins A & B, package insert. Meridian Bioscience, Inc. , Nice, France. http://www. meridianbios cience. com/Content/Assets/Files/2. 1%20%20C. %20difficile%20Product s/Package-Insert-ImmunoCard-Toxins-A-and-B. pdf.
42. Polage CR, Chin DL, Leslie JL, Tang J, Cohen SH, Solnick JV. 2012. Outcomes in patients tested for Clostridium difficile toxins. Diagn Microbiol Infect Dis 74:369-373. http://dx. doi. org/10. 1016/j. diagmicrobio . 2012. 08. 019.
43. Baker I, Leeming JP, Reynolds R, Ibrahim I, Darley E. 2013. Clinical relevance of a positive molecular test in the diagnosis of Clostridium difficile infection. J Hosp Infect 84:311-315. http://dx. doi. org/10. 1016/j. jhin . 2013. 05. 006.
44. Darkoh C, DuPont HL, Norris SJ, Kaplan HB. 2015. Toxin synthesis by Clostridium difficile is regulated through quorum signaling. mBio 6(2): e02569-14. http://dx. doi. org/10. 1128/mBio. 02569-14.
45. Tenover FC, Novak-Weekley S, Woods CW, Peterson LR, Davis T, Schreckenberger P, Fang FC, Dascal A, Gerding DN, Nomura JH, Goering RV, Akerlund T, Weissfeld AS, Baron EJ, Wong E, Marlowe EM, Whitmore J, Persing DH. 2010. Impact of strain type on detection of toxigenic Clostridium difficile: comparison of molecular diagnostic and enzyme immunoassay approaches. J Clin Microbiol 48:3719-3724. http: //dx. doi. org/10. 1128/JCM. 00427-10.
46. Antunes A, Dupuy B. 2010. Molecular methods to study transcriptional regulation of Clostridium difficile toxin genes. Methods Mol Biol 646:93-115. http://dx. doi. org/10. 1007/978-1-60327-365-7-7.
47. Karlsson S, Burman LG, Akerlund T. 2008. Induction of toxins in Clostridium difficile is associated with dramatic changes of its metabolism. Microbiology 154:3430-3436. http://dx. doi. org/10. 1099/mic. 0 . 2008/019778-0.
48. Kader HA, Piccoli DA, Jawad AF, McGowan KL, Maller ES. 1998. Single toxin detection is inadequate to diagnose Clostridium difficile diarrhea in pediatric patients. Gastroenterology 115:1329-1334. http://dx. doi. org/10 . 1016/S0016-5085(98)70009-5.
49. Riegler M, Sedivy R, Pothoulakis C, Hamilton G, Zacherl J, Bischof G, Cosentini E, Feil W, Schiessel R, LaMont JT. 1995. Clostridium difficile toxin B is more potent than toxin A in damaging human colonic epithelium in vitro. J Clin Invest 95:2004-2011. http://dx. doi. org/10. 1172 /JCI117885.