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Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification

  • Authors:
    • Lan Yu
    • Huan Li
    • Xiangna Zhao
    • Xuesong Wang
    • Xiao Wei
    • Weishi Lin
    • Puyuan Li
    • Lihong Cui
    • Jing Yuan
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, Navy General Hospital, Beijing 100048, P.R. China, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing 100071, P.R. China
    Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 4781-4788
    |
    Published online on: September 21, 2017
       https://doi.org/10.3892/etm.2017.5178
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Abstract

The binary toxin Clostridium difficile transferase (CDT) is frequently observed in C. difficile strains and is associated with an increased severity of C. difficile infection. CDT‑producing C. difficile infections cause higher fatality rates than infections with CDT negative isolates. Thus, the rapid and accurate identification of a CDT positive C. difficile infection is critical for effective treatment. The present study demonstrates how loop‑mediated isothermal amplification (LAMP) can be used to detect CDT‑producing C. difficile based on visual observation. This is a low complexity, rapid molecular method that has the potential to be used within a point of care setting. The specificity and sensitivity of the primers in the LAMP reactions for CDT detection were determined using two different methods, a real‑time turbidity monitor and visual detection after the addition of calcein to the reaction tube. The results revealed that target DNA was amplified and visualized by these two detection methods within 60 min at a temperature of 60˚C. The sensitivity of the LAMP assay was identified to be 10‑fold greater than that of polymerase chain reaction analysis. When 25 alternative bacterial strains lacking CDT were tested, the results of the amplification were negative, confirming the specificity of the primers. In conclusion, the visual LAMP method established in the present study may be a rapid, reliable and cost‑effective tool for detecting CDT‑producing C. difficile strains at the point of care.
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1 

Doosti A and Mokhtari-Farsani A: Study of the frequency of Clostridium difficile tcdA, tcdB, cdtA and cdtB genes in feces of Calves in south west of Iran. Ann Clin Microbiol Antimicrob. 13:212014. View Article : Google Scholar : PubMed/NCBI

2 

Norén T, Unemo M, Magnusson C, Eiserman M and Matussek A: Evaluation of the rapid loop-mediated isothermal amplification assay Illumigene for diagnosis of Clostridium difficile in an outbreak situation. APMIS. 122:155–160. 2014. View Article : Google Scholar : PubMed/NCBI

3 

Boyanton BL Jr, Sural P, Loomis CR, Pesta C, Gonzalez-Krellwitz L, Robinson-Dunn B and Riska P: Loop-mediated isothermal amplification compared to real-time PCR and enzyme immunoassay for toxigenic Clostridium difficile detection. J Clin Microbiol. 50:640–645. 2012. View Article : Google Scholar : PubMed/NCBI

4 

Kim J, Seo MR, Kang JO, Choi TY and Pai H: Clinical and Microbiologic characteristics of Clostridium difficile infection caused by binary toxin producing strain in Korea. Infect Chemother. 45:175–183. 2013. View Article : Google Scholar : PubMed/NCBI

5 

Papatheodorou P, Hornuss D, Nölke T, Hemmasi S, Castonguay J, Picchianti M and Aktories K: Clostridium difficile binary toxin CDT induces clustering of the lipolysis-stimulated lipoprotein receptor into lipid rafts. MBio. 4:e00244–e00213. 2013. View Article : Google Scholar : PubMed/NCBI

6 

Gerding DN, Johnson S, Rupnik M and Aktories K: Clostridium difficile binary toxin CDT: Mechanism, epidemiology, and potential clinical importance. Gut Microbes. 5:15–27. 2014. View Article : Google Scholar : PubMed/NCBI

7 

Silva RO, Santos RL, Pires PS, Pereira LC, Pereira ST, Duarte MC, de Assis RA and Lobato FC: Detection of toxins A/B and isolation of Clostridium difficile and Clostridium perfringens from dogs in Minas Gerais, Brazil. Braz J Microbiol. 44:133–137. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N and Hase T: Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28:E632000. View Article : Google Scholar : PubMed/NCBI

9 

Kim JB, Han AR, Park EY, Kim JY, Cho W, Lee J, Seo EK and Lee KT: Inhibition of LPS-induced iNOS, COX-2 and cytokines expression by poncirin through the NF-kappaB inactivation in RAW 264.7 macrophage cells. Biol Pharm Bull. 30:2345–2351. 2007. View Article : Google Scholar : PubMed/NCBI

10 

Hori S, Nomura T and Sakaguchi S: Control of regulatory T cell development by the transcription factor Foxp3. Science. 299:1057–1061. 2003. View Article : Google Scholar : PubMed/NCBI

11 

Gandelman O, Jackson R, Kiddle G and Tisi L: Loop-mediated amplification accelerated by stem primers. Int J Mol Sci. 12:9108–9124. 2011. View Article : Google Scholar : PubMed/NCBI

12 

Liu C, Jiang DN, Xiang GM, Luo FK, Liu LL, Yu JC and Pu XY: DNA detection of Clostridium difficile infection based on real-time resistance measurement. Genet Mol Res. 12:3296–3304. 2013. View Article : Google Scholar : PubMed/NCBI

13 

Mori Y, Nagamine K, Tomita N and Notomi T: Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun. 289:150–154. 2001. View Article : Google Scholar : PubMed/NCBI

14 

Gavin MA, Rasmussen JP, Fontenot JD, Vasta V, Manganiello VC, Beavo JA and Rudensky AY: Foxp3-dependent programme of regulatory T-cell differentiation. Nature. 445:771–775. 2007. View Article : Google Scholar : PubMed/NCBI

15 

Pollock NR: Ultrasensitive detection and quantification of toxins for optimized diagnosis of Clostridium difficile infection. J Clin Microbiol. 54:259–264. 2016. View Article : Google Scholar : PubMed/NCBI

16 

Hicke B, Pasko C, Groves B, Ager E, Corpuz M, Frech G, Munns D, Smith W, Warcup A, Denys G, et al: Automated detection of toxigenic Clostridium difficile in clinical samples: Isothermal tcdB amplification coupled to array-based detection. J Clin Microbiol. 50:2681–2687. 2012. View Article : Google Scholar : PubMed/NCBI

17 

Mansour SM, Ali H, Chase CC and Cepica A: Loop-mediated isothermal amplification for diagnosis of 18 World Organization for Animal Health (OIE) notifiable viral diseases of ruminants, swine and poultry. Anim Health Res Rev. 16:89–106. 2015. View Article : Google Scholar : PubMed/NCBI

18 

Yang L, Li J, Bi Y, Xu L and Liu W: Development and application of a reverse transcription loop-mediated isothermal amplification method for rapid detection of Duck hepatitis A virus type 1. Virus Genes. 45:585–589. 2012. View Article : Google Scholar : PubMed/NCBI

19 

Hirayama H, Kageyama S, Moriyasu S, Sawai K and Minamihashi A: Embryo sexing and sex chromosomal chimerism analysis by loop-mediated isothermal amplification in cattle and water buffaloes. J Reprod Dev. 59:321–326. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Seki M, Yamashita Y, Torigoe H, Tsuda H, Sato S and Maeno M: Loop-mediated isothermal amplification method targeting the lytA gene for detection of Streptococcus pneumoniae. J Clin Microbiol. 43:1581–1586. 2005. View Article : Google Scholar : PubMed/NCBI

21 

Kim DW, Kilgore PE, Kim EJ, Kim SA, Anh DD, Dong BQ, Kim JS and Seki M: The enhanced pneumococcal LAMP assay: A clinical tool for the diagnosis of meningitis due to Streptococcus pneumoniae. PLoS One. 7:e429542012. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Yu L, Li H, Zhao X, Wang X, Wei X, Lin W, Li P, Cui L and Yuan J: Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification. Exp Ther Med 14: 4781-4788, 2017.
APA
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W. ... Yuan, J. (2017). Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification. Experimental and Therapeutic Medicine, 14, 4781-4788. https://doi.org/10.3892/etm.2017.5178
MLA
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W., Li, P., Cui, L., Yuan, J."Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification". Experimental and Therapeutic Medicine 14.5 (2017): 4781-4788.
Chicago
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W., Li, P., Cui, L., Yuan, J."Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification". Experimental and Therapeutic Medicine 14, no. 5 (2017): 4781-4788. https://doi.org/10.3892/etm.2017.5178
Copy and paste a formatted citation
x
Spandidos Publications style
Yu L, Li H, Zhao X, Wang X, Wei X, Lin W, Li P, Cui L and Yuan J: Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification. Exp Ther Med 14: 4781-4788, 2017.
APA
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W. ... Yuan, J. (2017). Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification. Experimental and Therapeutic Medicine, 14, 4781-4788. https://doi.org/10.3892/etm.2017.5178
MLA
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W., Li, P., Cui, L., Yuan, J."Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification". Experimental and Therapeutic Medicine 14.5 (2017): 4781-4788.
Chicago
Yu, L., Li, H., Zhao, X., Wang, X., Wei, X., Lin, W., Li, P., Cui, L., Yuan, J."Rapid visual detection of binary toxin producing Clostridium difficile by loop‑mediated isothermal amplification". Experimental and Therapeutic Medicine 14, no. 5 (2017): 4781-4788. https://doi.org/10.3892/etm.2017.5178
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