›› 2017, Vol. 44 ›› Issue (11): 3334-3339.doi: 10.16431/j.cnki.1671-7236.2017.11.032

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Establishment and Application of Visual RT-LAMP for Rapid Detection of Duck Tembusu Virus

WU Zhi1, XIA Wen-long1, CAI Shu-dong1, GUO Chang-ming1, YUAN Wei-feng2, WANG Yong-juan1   

  1. 1. Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals, Jiangsu Animal Husbandry and Veterinary College, Taizhou 225300, China;
    2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2017-07-03 Online:2017-11-20 Published:2017-11-21

Abstract:

To realize the rapid detection of duck tembusu virus (DTMUV), a set of specific primers was designed according to the highly conserved sequence of DTMUV E protein gene published in GenBank. A sensitive and convenient RT-LAMP amplification method was established by optimizing the concentrations of each component, reaction time and temperature in the reaction system. The results showed that the influence of betaine on the reaction system was not obvious under the optimal conditions. This method was 100 times more sensitive than the conventional RT-PCR. The RT-LAMP method specifically amplified tembusu virus but not other common avian viruses. The results could be differentiated by naked eyes. 74 clinic suspicious samples were tested and 65 of them were positive. These results suggested that the RT-LAMP assay could be used as a method for the diagnosis and detection of clinical cases, and molecular epidemiological investigation of duck tembusu virus disease.

Key words: duck tembusu virus (DTMUV); RT-LAMP; visualization; rapid detection

CLC Number: