A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout ( Oncorhynchus mykiss )

A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic... A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout ( Oncorhynchus mykiss ). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 °C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a10-fold higher sensitivity was seen for the LAMP protocols. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout ( Oncorhynchus mykiss )

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Publisher
Springer-Verlag
Copyright
Copyright © 2005 by Springer-Verlag/Wien
Subject
Biomedicine; Medical Microbiology; Virology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-004-0468-7
Publisher site
See Article on Publisher Site

Abstract

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout ( Oncorhynchus mykiss ). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 °C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a10-fold higher sensitivity was seen for the LAMP protocols.

Journal

Archives of VirologySpringer Journals

Published: May 1, 2005

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