A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda

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

doi:10.1007/s00343-012-1244-2

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A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda

Updated：2023-05-16

- A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda
- Journal of Oceanology and Limnology Vol. 30, Issue 5, Pages: 731-737(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/s00343-012-1244-2
  CLC：
- Online First：05 September 2012，
  
  Published：2012
- Accepted：
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Xie, G., Huang, J., Zhang, Q. et al. A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda, ., Chin. J. Ocean. Limnol. 30, 731–737 (2012). https://doi.org/10.1007/s00343-012-1244-2
DOI：

Xie, G., Huang, J., Zhang, Q. et al. A real-time PCR targeted to the upstream regions of HlyB for specific detection of Edwardsiella tarda, ., Chin. J. Ocean. Limnol. 30, 731–737 (2012). https://doi.org/10.1007/s00343-012-1244-2 DOI：

摘要

Abstract

Edwardsiella tarda has become one of the most important emerging pathogens in aquaculture industry. Therefore

a rapid

reproducible

and sensitive method for detection and quantification of this pathogen is needed urgently. To achieve this purpose

we developed a TaqMan-based real-time PCR assay for detection and quantification of E. tarda. The assay targets the hemolysin activator HlyB domain protein of E. tarda. Our optimized TaqMan assay is capable of detecting as little as 40 fg of genomic DNA per reaction. A standard curve was generated from the threshold cycle values (y) against log10 (E. tarda genomic DNA concentration) as x. The intra- and inter-assay coefficient of variation (CV) values were less than 2.06% and 1.05% respectively

indicating that the assay had good reproducibility. This method is highly specific to E. tarda strains

as it shows no cross-reactivity to Edwardsiella ictaluri

a member of the same genus

or to nine other fish-pathogenic bacteria species belonging to three other genera. This sensitive and specific real-time PCR assay provides a valuable tool for diagnostic quantitation of E. tarda in clinical samples.

关键词

Keywords

references

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