An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene

Guosi Xie; Qingli Zhang; Nana Han; Chengyin Shi; Xiuhua Wang; Qinghui Liu; Jie Huang

doi:10.1007/s00343-012-1293-6

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An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene

Updated：2023-05-16

- An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene
- Journal of Oceanology and Limnology Vol. 30, Issue 4, Pages: 595-603(2012)
- Affiliations：
  
  1. Key Laboratory of Marine Fishery Resources Sustainable Utilization Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences
  2. College of Fisheries and Life Science Shanghai Ocean University
- Author bio：
- Funds：
- DOI：10.1007/s00343-012-1293-6
  CLC：
- Online First：02 July 2012，
  
  Published：2012
- Accepted：
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Xie, G., Zhang, Q., Han, N. et al. An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene., Chin. J. Ocean. Limnol. 30, 595–603 (2012). https://doi.org/10.1007/s00343-012-1293-6
DOI：

Xie, G., Zhang, Q., Han, N. et al. An improved method for detection of Edwardsiella tarda by loop-mediated isothermal amplification by targeting the EsrB gene., Chin. J. Ocean. Limnol. 30, 595–603 (2012). https://doi.org/10.1007/s00343-012-1293-6 DOI：

摘要

Abstract

Edwardsiella tarda is a major pathogen in aquatic environments that can cause heavy economic losses. An improved method for quick and accurate detection of E. tarda by loop-mediated isothermal amplification (LAMP) with two additional loop primers was developed by targeting the EsrB gene (EsrB — LAMP). In this method

the Mg2+ concentration

reaction temperature

and reaction time were optimized to 8 mmol/L

61°C

and 40 min

respectively. The detection limit with the EsrB gene was as low as 10 copies

which is 100 times more sensitive than that of conventional polymerase chain reaction (PCR). The EsrB-LAMP assay was shown more sensitive and rapid than previously reported LAMP assays targeting the hemolysin gene ( hemolysin -LAMP) for detection of E. tarda. The EsrB -LAMP was also highly specific to E. tarda and had no cross-reaction with 13 other strains of bacteria. The assay can be carried out in a simple heating device and the EsrB-LAMP products can be visually detected by adding fluorescent dye to the reaction mixture. Taken together

the improved EsrB-LAMP diagnostic protocol has the potential for detection of E. tarda from indoor and outdoor samples.

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Keywords

references

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