Detection of red-spotted grouper nervous necrosis virus by loop-mediated isothermal amplification.
Red-spotted grouper nervous necrosis virus (RGNNV) causes high mortality in marine fish larvae cultured in China. To control better an outbreak of this virus, a rapid, specific and sensitive detection method based on loop-mediated isothermal amplification (LAMP) was developed. A set of four primers, two outer and two inner, was designed from RGNNV genome RNA. The LAMP reaction mix was optimized. The method was specific as no cross-reaction was observed between white spot syndrome virus, koi herpesvirus, infectious spleen and kidney necrosis virus, mud crab reovirus, and grass carp hemorrhage virus. The sensitivity of LAMP was 100-fold higher than the nested PCR in detecting the presence of RGNNV. RGNNV was detected in the brain of Trachinotus ovatus that showed typical symptoms of NNV infection, with the standardized LAMP procedure.
Xu
Hai-Dong
HD
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 231 Xingang Road West, Guangzhou 510300, China.
Feng
Juan
J
Guo
Zhi-Xun
ZX
Ou
You-Jun
YJ
Wang
Jiang-Yong
JY
D000818:Animals; D001483:Base Sequence; D001622:Betaine / Q000378:metabolism; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology*; D005399:Fishes / Q000821:virology*; D008274:Magnesium / Q000378:metabolism; D008969:Molecular Sequence Data; D030381:Nodaviridae / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D001622:Betaine; D008274:Magnesium