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《Acta Agriculturae Zhejiangensis》 2014-01
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Development and evaluation of RT-LAMP for the rapid detection of goose-origin Tembusu virus

HAN Kai-kai;LI Yin;HUANG Xin-mei;ZHAO Dong-min;LIU Yu-zhuo;XIE Xing-xing;YOU Yuan;Key Laboratory of Veterinary Biologicals engineering and technology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences;  
In this study,a RT-LAMP( reverse transcription loop-mediated isothermal amplification) assay was developed to rapidly detect and diagnose goose tembusu virus( GTMUV). Using Primer Explorer V4 software,a series of specific primers were designed based on the nucleotide sequences of GTMUV NS5 gene available in GenBank. These primers targeted 6 distinct conserved sequences according to the conserved region of NS5 gene. The results showed the RT-LAMP method specifically amplified GTMUV but not other common avian disease RNA viruses,and the optimum reaction temperature and time were verified to be 60℃ and 60 min. Compared with other methods,this method was 100 times more sensitive than the conventional PCR. Meanwhile,it did not require complicated thermal cycling steps and denaturation,with the addition of SYBR GreenⅠ dye in the reaction mixture,and the results could be differentiated by naked eyes. Thus,the results proved that RT-LAMP was a reliable,accurate,simple and practical method for the detection of avian Tembusu virus in the field.
【Fund】: 国家自然科学基金项目(31172345);; 江苏省农业科技自主创新资金项目[cx(12)5048]
【CateGory Index】: S858.33
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