title: Reverse transcriptase loop-mediated isothermal amplification assay for infectious hematopoietic necrosis virus in Oncorhynchus keta.
abstract: A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting infectious hematopoietic necrosis virus (IHNV) from chum salmon Oncorhynchus keta in South Korea with high specificity, sensitivity and rapidity. A set of 6 IHNV-specific primers was designed, based on the G-protein sequence of IHNV (PRT strain), recognizing 8 distinct sequences of the target RNA. The assay was optimized to detect IHNV at 63 degrees C for 30 min. The limit of detection was 0.01 fg of RNA extracted from IHNV-infected CHSE-214 cells, compared with 1.0 fg for nested RT-PCR. The applicability of this RT-LAMP assay was further tested by comparison with nested RT-PCR using field samples. Of 473 samples tested, 191 samples (40.38%) were IHNV-positive by RT-LAMP, whereas 162 samples (34.25%) were IHNV-positive by nested RT-PCR. These results indicate that, because of its high sensitivity and rapidity, the RT-LAMP assay is useful for early diagnosis of IHN.
authors.affiliation: Faculty of Marine Bioscience & Technology, Gangneung-Wonju National University, Gangneung 210-702, South Korea.
authors.forename: Rungkarn
authors.initials: R
authors.lastname: Suebsing
authors.forename: Chan-Hyeok
authors.initials: CH
authors.lastname: Jeon
authors.forename: Myung-Joo
authors.initials: MJ
authors.lastname: Oh
authors.forename: Jeong-Ho
authors.initials: JH
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D029142:Infectious hematopoietic necrosis virus; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D017685:Oncorhynchus keta; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018353:Rhabdoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity