›› 2018, Vol. 45 ›› Issue (1): 22-31.doi: 10.16431/j.cnki.1671-7236.2018.01.003

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Establishment of RT-LAMP Visual Detection Method for Goose Newcastle Disease Virus

LIU Wenjun1, HUANG Yunmao1, YANG Youtian2, XU Danning1, CAO Nan1, ZHOU Derong1, TIAN Yunbo1   

  1. 1. Key Laboratory of Waterfowl Healthy Breeding in Guangdong, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China;
    2. Foshan University, Foshan 528000, China
  • Received:2017-06-13 Online:2018-01-20 Published:2018-01-20

Abstract:

This study was aimed to establish a Real-time and easy-to-diagnose method for the detection of goose Newcastle disease virus (GNDV) at the grassroots level. Based on the highly homologous conserved sequence of GNDV F gene published in GenBank, specific primers were designed and the calcein/manganese chloride indicator was added to the reaction system to establish a visual RT-LAMP detection method. The results showed that the detection method could specifically detect GNDV and NDV Lasota vaccine strains, and other viruses such as GPV, AIV and DTMUV were negative. The minimum detection limit of RNA was 10 pg. The reaction procedure did not require complex instruments such as PCR instrument, and the reaction was completed within 50 min. The results were observed by the naked eye. This method was specificity, high sensitivity, and safe operation, simple and fast, could meet the primary screening of GNDV.

Key words: goose Newcastle disease virus (GNDV); loop mediated isothermal amplification (LAMP); calcein

CLC Number: