DAO 94:1-8 (2011)  -  DOI: https://doi.org/10.3354/dao02310

Reverse transcriptase loop-mediated isothermal amplification assay for infectious hematopoietic necrosis virus in Oncorhynchus keta

Rungkarn Suebsing1, Chan-Hyeok Jeon1, Myung-Joo Oh2, Jeong-Ho Kim1,*

1Faculty of Marine Bioscience & Technology, Gangneung-Wonju National University, Gangneung 210-702, South Korea
2Department of Aqualife Medicine, Chonnam National University, Yosu 550-749, South Korea
*Corresponding author. Email:

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°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.


KEY WORDS: RT-LAMP · IHNV · Chum salmon · Oncorhynchus keta


Full text in pdf format 
Cite this article as: Suebsing R, Jeon CH, Oh MJ, Kim JH (2011) Reverse transcriptase loop-mediated isothermal amplification assay for infectious hematopoietic necrosis virus in Oncorhynchus keta. Dis Aquat Org 94:1-8. https://doi.org/10.3354/dao02310

Export citation
Mail this link - Contents Mailing Lists - RSS
- -