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Volume 37 Issue 4
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Article Navigation >  Acta Oceanologica Sinica  > 2018  >  37(4): 83-88
ZHANG Lina, WANG Mingyi, CONG Dianxia, DING Shuyan, CONG Rinan, YUE Jinyong, GENG Jianli, HU Chengjin. Rapid, specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene[J]. Acta Oceanologica Sinica, 2018, 37(4): 83-88. doi: 10.1007/s13131-018-1182-8
Citation: ZHANG Lina, WANG Mingyi, CONG Dianxia, DING Shuyan, CONG Rinan, YUE Jinyong, GENG Jianli, HU Chengjin. Rapid, specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene[J]. Acta Oceanologica Sinica, 2018, 37(4): 83-88. doi: 10.1007/s13131-018-1182-8
shu

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

doi: 10.1007/s13131-018-1182-8
  • 1.

    Department of Laboratory Medicine, General Hospital of Jinan Military Region of PLA, Jinan 250031, China

  • 2.

    Department of Laboratory Medicine, General Hospital of Jinan Military Region of PLA, Jinan 250031, China;Department of Central Laboratory, Weihai Municipal Hospital affiliated to Dalian Medical University, Weihai 264200, China

  • 3.

    Department of Central Laboratory, Weihai Municipal Hospital affiliated to Dalian Medical University, Weihai 264200, China

  • Received Date: 2017-03-27
    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℃ 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.
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