LAMP for the rapid diagnosis of iridovirus in aquaculture  

作　　者：Yepin Yu Zituo Yang Le Wang Fei Sun May Lee Yanfei Wen Qiwei Qin Gen Hua Yue 

机构地区：[1]Temasek Life Sciences Laboratory,National University of Singapore,1 Research Link,117604,Singapore [2]Joint Laboratory of Guangdong Province and Hong Kong Region on Marine Bioresource Conservation and Exploitation,College of Marine Sciences,South China Agricultural University,Guangzhou,510642,China [3]Department of Biological Sciences,National University of Singapore,14 Science Drive,117543,Singapore

出　　处：《Aquaculture and Fisheries》2022年第2期158-165,共8页渔业学报（英文）

基　　金：This study was supported by the National Natural Science Foundation of China(41806161);the internal funds of Temasek Life Sciences Lab,Singapore.

摘　　要：Iridoviruses are DNA virus and have caused huge economic losses in the aquaculture industry.The aim of this study was to establish a colorimetric loop-mediated isothermal amplification(LAMP)protocol for the on-site detection of Singapore grouper iridovirus(SGIV).The SGIV-VP61 gene was chosen as the target gene to develop a colorimetric LAMP assay.The optimized condition of the colorimetric LAMP assay was incubation at 63℃ for 1 h.Samples infected with SGIV could be detected with the color change from yellow into pink.The sensitivity of the developed assay is 5.66 copies/μL of the viral DNA template.This sensitivity was about 1000 times higher than that of conventional PCR while it was slightly lower than the one-step semi-nested PCR assay.A total of 60 DNA samples extracted from the fin tissue of the SGIV-infected Asian seabass were examined for SGIV by colorimetric LAMP,semi-nested PCR and conventional PCR.The results of the colorimetric LAMP assay showed 94.87%agreement with the semi-nested PCR.In addition,the DNA extraction method using NaOH showed a better performance in the colorimetric LAMP assay.Taken together,the colorimetric LAMP established was a sensitive,rapid and specific method for the detection of SGIV.SGIV was not detected in samples randomly taken from a genetically improved line of the Asian seabass.However,some seabass obtained from the local markets were found to contain SGIV.Thus,the LAMP assay has the potential application in the diagnosis of iridovirus diseases in the aquaculture industry.

关 键 词：FISH DISEASE IRIDOVIRUS Diagnosis PCR SEABASS 

分 类 号：S94[农业科学—水产养殖]