Acta Oceanologica Sinica

, Volume 37, Issue 4, pp 83–88 | Cite as

Rapid, specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene

  • Lina Zhang
  • Mingyi Wang
  • Dianxia Cong
  • Shuyan Ding
  • Rinan Cong
  • Jinyong Yue
  • Jianli Geng
  • Chengjin Hu
Article
  • 5 Downloads

Abstract

Vibrio vulnificus is an estuarine bacterial pathogen for human. The rapid, specific and sensitive detection of V. vulnificus is urgently needed for early disease diagnosis and timely treatment of V. vulnificus infection. In the study, a loop-mediated isothermal amplification (LAMP) technique was developed for V. vulnificus detection with a set of primers, composed of two out primers and two inner primers targeted to vvhA gene. The optimal amplification temperature was 63°C and the reaction only took 35 min. The amplification products could not only be detected by agarose gel electrophoresis with ladder-like pattern bands, but also could be visualized using calcein with naked eye directly. Forty-five strains were tested for the specificity of LAMP assay, and all the V. vulnificus strains were identified correctly while other strains were negative results. The sensitive of the new LAMP assay was 100-fold more sensitive than the conventional PCR. Meanwhile, all the V. vulnificus strains were detected correctly in spiked, clinical and environmental samples by the new LAMP assay. Compared with other well-known techniques, the new LAMP assay targeted to vvhA gene was extremely rapid, simple, sensitive and specific for V. vulnificus identification.

Keywords

Vibrio vulnificus loop-mediated isothermal amplification vvhA gene 

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Copyright information

© The Chinese Society of Oceanography and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Lina Zhang
    • 1
  • Mingyi Wang
    • 1
    • 2
  • Dianxia Cong
    • 2
  • Shuyan Ding
    • 2
  • Rinan Cong
    • 2
  • Jinyong Yue
    • 2
  • Jianli Geng
    • 2
  • Chengjin Hu
    • 1
  1. 1.Department of Laboratory MedicineGeneral Hospital of Jinan Military Region of PLAJinanChina
  2. 2.Department of Central LaboratoryWeihai Municipal Hospital affiliated to Dalian Medical UniversityWeihaiChina

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