Intest Res.  2018 Jan;16(1):109-115. 10.5217/ir.2018.16.1.109.

Rapid and accurate diagnosis of Clostridium difficile infection by real-time polymerase chain reaction

• Song PH
• Min JH
• Kim YS
• Jo SY
• Kim EJ
• Lee KJ
• Lee J
• Sung H
• Moon JS
• Whang DH

• KMID: 2402654
• DOI: http://doi.org/10.5217/ir.2018.16.1.109

Abstract

BACKGROUND/AIMS
The incidence and severity of Clostridium difficile infection (CDI) have increased worldwide, resulting in a need for rapid and accurate diagnostic methods.
METHODS
A retrospective study was conducted to compare CDI diagnosis methods between January 2014 and December 2014. The stool samples, which were obtained in presumptive CDI patients, were compared for their diagnostic accuracy and rapidity, including real-time polymerase chain reaction (PCR) of toxin genes, C. difficile toxin assay, and culture for C. difficile.
RESULTS
A total of 207 cases from 116 patients were enrolled in this study and 117 cases (56.5%) were diagnosed as having CDI. Among the 117 cases, the sensitivities of real-time PCR, C. difficile toxin assay, and culture for C. difficile were 87.2% (102 cases; 95% CI, 80.7%-92.8%), 48.7% (57 cases; 95% CI, 41.0%-59.8%), and 65.0% (76 cases; 95% CI, 60.2%-78.5%), respectively (P < 0.005). Notably, 34 cases (29.0%) were diagnosed with CDI by real-time PCR only. The time required to obtain results was 2.27 hours (136.62±82.51 minutes) for real-time PCR, 83.67 hours (5,020.66±3,816.38 minutes) for toxin assay, and 105.79 hours (6,347.68±3,331.46 minutes) for culture (P < 0.005), respectively.
CONCLUSIONS
We confirmed that real-time PCR of toxin genes is the most effective diagnostic method for accurate and early diagnosis of CDI. It also helps to diagnose hypervirulent CDI, such as ribotype 027 infection.

Keyword

Clostridium difficile infection; Polymerase chain reaction; Diagnostic method

MeSH Terms

Clostridium difficile*
Clostridium*
Diagnosis*
Early Diagnosis
Humans
Incidence
Methods
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction*
Retrospective Studies
Ribotyping

Figure

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  Fig. 1 Study process. A total of 207 cases from 116 patients were initially enrolled in this study and from these, 117 cases from 90 patients were diagnosed as Clostridium difficile infection (CDI).

Reference

1. O'Connor JR, Johnson S, Gerding DN. Clostridium difficile infection caused by the epidemic BI/NAP1/027 strain. Gastroenterology. 2009; 136:1913–1924. PMID: 19457419.

2. Pépin J, Valiquette L, Alary ME, et al. Clostridium difficile-associated diarrhea in a region of Quebec from 1991 to 2003: a changing pattern of disease severity. CMAJ. 2004; 171:466–472. PMID: 15337727.

3. Ricciardi R, Rothenberger DA, Madoff RD, Baxter NN. Increasing prevalence and severity of Clostridium difficile colitis in hospitalized patients in the United States. Arch Surg. 2007; 142:624–631. PMID: 17638799.

4. Burke KE, Lamont JT. Clostridium difficile infection: a worldwide disease. Gut Liver. 2014; 8:1–6. PMID: 24516694.

5. Kim YS, Han DS, Kim YH, et al. Incidence and clinical features of Clostridium difficile infection in Korea: a nationwide study. Epidemiol Infect. 2013; 141:189–194. PMID: 22717061.

6. Doh YS, Kim YS, Jung HJ, et al. Long-term clinical outcome of Clostridium difficile infection in hospitalized patients: a single center study. Intest Res. 2014; 12:299–305. PMID: 25374496.

7. Yoon SY, Jung SA, Na SK, et al. What's the clinical features of colitis in elderly people in long-term care facilities? Intest Res. 2015; 13:128–134. PMID: 25931997.

8. Mueller S, Saunier K, Hanisch C, et al. Differences in fecal microbiota in different European study populations in relation to age, gender, and country: a cross-sectional study. Appl Environ Microbiol. 2006; 72:1027–1033. PMID: 16461645.

9. Barbut F, Monot M, Rousseau A, et al. Rapid diagnosis of Clostridium difficile infection by multiplex real-time PCR. Eur J Clin Microbiol Infect Dis. 2011; 30:1279–1285. PMID: 21487764.

10. Berry N, Sewell B, Jafri S, et al. Real-time polymerase chain reaction correlates well with clinical diagnosis of Clostridium difficile infection. J Hosp Infect. 2014; 87:109–114. PMID: 24795170.

11. de Jong E, de Jong AS, Bartels CJ, van der Rijt-van den Biggelaar C, Melchers WJ, Sturm PD. Clinical and laboratory evaluation of a real-time PCR for Clostridium difficile toxin A and B genes. Eur J Clin Microbiol Infect Dis. 2012; 31:2219–2225. PMID: 22327373.

12. Poutanen SM, Simor AE. Clostridium difficile-associated diarrhea in adults. CMAJ. 2006; 171:51–58.

13. Sewell B, Rees E, Thomas I, Ch’ng CL, Isaac M, Berry N. Cost and impact on patient length of stay of rapid molecular testing for Clostridium difficile. Infect Dis Ther. 2014; 3:281–293. PMID: 25183400.

14. Vaishnavi C. Clinical spectrum & pathogenesis of Clostridium difficile associated diseases. Indian J Med Res. 2010; 131:487–499. PMID: 20424299.

15. Goudarzi M, Seyedjavadi SS, Goudarzi H, Mehdizadeh Aghdam E, Nazeri S. Clostridium difficile infection: epidemiology, pathogenesis, risk factors, and therapeutic options. Scientifica (Cairo). 2014; 2014:916826. PMID: 24991448.

16. Warny M, Pepin J, Fang A, et al. Toxin production by an emerging strain of Clostridium difficile associated with outbreaks of severe disease in North America and Europe. Lancet. 2005; 366:1079–1084. PMID: 16182895.

17. Tae CH, Jung SA, Song HJ, et al. The first case of antibiotic-associated colitis by Clostridium difficile PCR ribotype 027 in Korea. J Korean Med Sci. 2009; 24:520–524. PMID: 19543521.

18. Kim J, Kang JO, Kim H, et al. Epidemiology of Clostridium difficile infections in a tertiary-care hospital in Korea. Clin Microbiol Infect. 2013; 19:521–527. PMID: 22712697.

19. Kim H, Lee Y, Moon HW, Lim CS, Lee K, Chong Y. Emergence of Clostridium difficile ribotype 027 in Korea. Korean J Lab Med. 2011; 31:191–196. PMID: 21779194.

20. Peppard WJ, Ledeboer NA. Implementation of polymerase chain reaction to rule out Clostridium difficile infection is associated with reduced empiric antibiotic duration of therapy. Hosp Pharm. 2014; 49:639–643. PMID: 25477583.

21. Catanzaro M, Cirone J. Real-time polymerase chain reaction testing for Clostridium difficile reduces isolation time and improves patient management in a small community hospital. Am J Infect Control. 2012; 40:663–666. PMID: 22153847.