Open Access

Evaluation of loop‑mediated isothermal amplification assays for rapid detection of blaKPC producing Serratia spp. in clinical specimens: A prospective diagnostic accuracy study

  • Authors:
    • Xinwei Liu
    • Dayang Zou
    • Chunxia Wang
    • Xiaoqian Zhang
    • Dongxu Pei
    • Wei Liu
    • Yongwei Li
  • View Affiliations

  • Published online on: February 1, 2021     https://doi.org/10.3892/etm.2021.9739
  • Article Number: 308
  • Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The prevalence of carbapenem‑resistant Serratia spp. is increasing owing to the propagation of β lactamase Klebsiella pneumoniae carbapenemase (blaKPC) and it has become one of the major global health concerns. As effective therapies for such resistant pathogens are limited, there is a great need for the rapid and sensitive characterization of the pathogen. In the present study, a loop‑mediated isothermal amplification (LAMP) method for the rapid detection of Serratia spp. with blaKPC in pure cultures and clinical specimens was developed. A calcein indicator and real‑time turbidity recording system were used to assess the LAMP reaction. The LAMP assay was compared with conventional PCR and real‑time PCR kits for the target pathogen. The desired amplification was achieved using selected primers and detection was possible using both the calcein indicator method and the real‑time turbity recording system at 65˚C for 60 min. The sensitivity of the detection system for blaKPC‑producing Serratia spp. reached a detection limit of 3.92 pg/µl DNA, which was 10 times more sensitive than conventional PCR. Specificity testing indicated that the primers were highly specific. Compared with conventional culture methods and real‑time PCR, the LAMP assay was more sensitive, easier for laboratory staff to master and less influenced by the clinical specimen matrix. In conclusion, a LAMP assay for blaKPC‑producing Serratia spp. that permitted rapid, sensitive and economical detection for this pathogen was successfully developed. Comparisons with alternative methods indicated that the LAMP assay was more feasible in a clinical setting.
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April-2021
Volume 21 Issue 4

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

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Spandidos Publications style
Liu X, Zou D, Wang C, Zhang X, Pei D, Liu W and Li Y: Evaluation of loop‑mediated isothermal amplification assays for rapid detection of <em>bla</em>KPC producing <em>Serratia spp.</em> in clinical specimens: A prospective diagnostic accuracy study. Exp Ther Med 21: 308, 2021
APA
Liu, X., Zou, D., Wang, C., Zhang, X., Pei, D., Liu, W., & Li, Y. (2021). Evaluation of loop‑mediated isothermal amplification assays for rapid detection of <em>bla</em>KPC producing <em>Serratia spp.</em> in clinical specimens: A prospective diagnostic accuracy study. Experimental and Therapeutic Medicine, 21, 308. https://doi.org/10.3892/etm.2021.9739
MLA
Liu, X., Zou, D., Wang, C., Zhang, X., Pei, D., Liu, W., Li, Y."Evaluation of loop‑mediated isothermal amplification assays for rapid detection of <em>bla</em>KPC producing <em>Serratia spp.</em> in clinical specimens: A prospective diagnostic accuracy study". Experimental and Therapeutic Medicine 21.4 (2021): 308.
Chicago
Liu, X., Zou, D., Wang, C., Zhang, X., Pei, D., Liu, W., Li, Y."Evaluation of loop‑mediated isothermal amplification assays for rapid detection of <em>bla</em>KPC producing <em>Serratia spp.</em> in clinical specimens: A prospective diagnostic accuracy study". Experimental and Therapeutic Medicine 21, no. 4 (2021): 308. https://doi.org/10.3892/etm.2021.9739