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Comparison of Three Terminal Detection Methods Based on Loop Mediated Isothermal Amplification (LAMP) Assay for Spring Viremia of Carp Virus (SVCV)

文献类型： 外文期刊

作者： Liu, Lu 1 ; Xu, Yuyan 2 ; Zhong, Weifang 1 ; Li, Lina 1 ; Li, Weizhe 1 ; Xiao, Qin 1 ;

作者机构： 1.Hebei Agr Univ, Coll Ocean, Qinhuangdao 066003, Hebei, Peoples R China

2.Minist Agr, Key Lab Control Qual & Safety Aquat Prod, Beijing 100141, Peoples R China

3.Chinese Acad Fishery Sci, Beijing 100141, Peoples R China

关键词： Agarose gel electrophoresis; Lateral flow dipstick (LFD); Loop-mediated isothermal amplification (LAMP); Spring viremia of carp virus (SVCV); SYBR Green I

期刊名称：TURKISH JOURNAL OF FISHERIES AND AQUATIC SCIENCES （ 影响因子：1.264； 五年影响因子：1.268 ）

ISSN： 1303-2712

年卷期： 2019 年 19 卷 9 期

页码：

收录情况： SCI

摘要： Spring viremia of carp virus (SVCV) is a significant cyprinid-pathogenic virus. SVCV detection is usually performed in a laboratory with apparatus. But the virus outbreaks are generally in fishery banks. The study was developed a loop-mediated isothermal amplification (LAMP) assay, and compared to three different terminal detection methods in order to achieve SVCV field-based detection. A set of six specific primers for LAMP were designed based on SVCV glycoprotein (G) gene. The reaction parameters of LAMP were optimized, and three terminal detection methods (SYBR Green I staining, lateral flow dipstick (LFD), and agarose gel electrophoresis) were applied respectively to the detection of LAMP products. The results showed that 8 mM Mg2+, 320 U/mL Bst DNA polymerase, 1.4 mM dNTP, and 1 M Betaine were optimum at 63 degrees C for 40 min. Among the three terminal ways, LFD was preferred for detection of LAMP products by comparing their sensitivity and specificity. The detection limit of LAMP combined LFD (LAMP-LFD) was 860 fg, and no cross-reaction with other aquaculture viruses. Thus, the presented LAMP-LFD was suitable for field-based detection of SVCV with its advantages of speed, simplicity, and disposability. Meanwhile, the study also provides a valuable alternative to immunoassays and PCR-based tests for other virus or bacteria.

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