Open Access

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

  • Published online on: September 21, 2017     https://doi.org/10.3892/etm.2017.5178
  • Pages: 4781-4788
  • Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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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November-2017
Volume 14 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

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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