title: A one-step molecular biology method for simple and rapid detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 strain.
abstract: Six reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primers designed against conserved regions of segment 6 (s6) gene were used for the detection of grass carp Ctenopharyngodon idella reovirus (GCRV) HZ08 subtype. The entire amplification could be completed within 40 min at 62·3° C. The RT-LAMP showed higher sensitivity than reverse-transcription polymerase chain reaction (RT-PCR). The RNA detection limit was 10 copies µl⁻¹ for RT-LAMP assay and 100 copies µl⁻¹ for conventional RT-PCR. In specificity tests, no cross-reactivity was detected in other viruses from common aquatic animals. In addition, the reaction results can be visualized by using calcein fluorescent dye. Furthermore, a total of 86 samples were tested by RT-LAMP, RT-PCR and virus isolation. The results demonstrated that all 54 specimens identified as positive by virus isolation were also positive when detected by RT-LAMP. Seven out of 54 samples, however, were misidentified by RT-PCR. The RT-LAMP method is more accurate than conventional RT-PCR. The results indicate that RT-LAMP has potential as a simple and rapid diagnosis technique for the detection of GCRV HZ08 subtype infection.
authors: Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China. zww8810303@163.com
authors: W W
authors: WW
authors: Zeng
authors: Q
authors: Q
authors: Wang
authors: Y Y
authors: YY
authors: Wang
authors: D H
authors: DH
authors: Xu
authors: S Q
authors: SQ
authors: Wu
mesh_terms: D000818:Animals; D002347:Carps / Q000821:virology*; D002460:Cell Line; D007668:Kidney / Q000166:cytology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012087:Reoviridae / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014776:Virus Cultivation / Q000379:methods