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Development and comparison of loop mediated isothermal amplification assay and polymerase chain reaction based on major capsid protein gene for detection of CyHV-2 infection in goldfish Carassius auratus (L.): LAMP and PCR for detection of CyHV-2 infection

Indian Agricultural Research Journals

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Title Development and comparison of loop mediated isothermal amplification assay and polymerase chain reaction based on major capsid protein gene for detection of CyHV-2 infection in goldfish Carassius auratus (L.): LAMP and PCR for detection of CyHV-2 infection
 
Creator Arathi Dharmaratnam
Raj Kumar
M. A. Pradeep
Neeraj Sood
T. Raja Swaminathan
 
Subject Goldfish
Cyprinid herpesvirus-2 (CyHV-2)
Diagnostic assays
PCR
LAMP assay
MCP gene
 
Description Cyprinid herpesvirus 2 (CyHV-2) is the etiological agent of goldfish herpesviral haematopoietic necrosis (GHVHN) that caused high economic losses in goldfish aquaculture. In this study, a loop-mediated isothermal amplification (LAMP) assay as well as a polymerase chain reaction (PCR) targeting major capsid protein (MCP) gene of CyHV-2 were standardised and evaluated for detection of CyHV-2. CyHV-2 was purified from infected fantail goldfish fin (FtGF) cells using ultracentrifugation and used as template for developing the diagnostic assays. The new LAMP and PCR assays are highly specific and did not amplify the nucleic acids of other fish pathogens tested, namely spring viraemia of carp virus (SVCV), Cyprinid herpesvirus-3 (CyHV-3), and viral nervous necrosis virus (VNNV), Aeromonas hydrophila, A. veronii, A. caviae, Edwardsiella tarda, Vibrio anguillarum, V. parahaemolyticus, V. harveyi and Proteus hauseri. Among the two assays developed, LAMP was found to be more sensitive, capable of detecting 10 copies of the plasmid construct containing 942 bp fragment of MCP gene of CyHV-2, while PCR could detect only 100 copies. The LAMP assay developed is a simple, rapid and reliable method for detection of CyHV-2 infection which can also be used in field conditions.
Keywords: Cyprinid herpesvirus-2 (CyHV-2), Diagnostic assays, Goldfish, LAMP assay, MCP gene, PCR
 
Publisher Central Marine Fisheries Research Institute (on behalf of Indian Council of Agricultural
 
Date 2024-06-30
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Peer-reviewed Article
 
Format application/pdf
 
Identifier https://epubs.icar.org.in/index.php/IJF/article/view/127966
10.21077/ijf.2024.71.2.127966-11
 
Source Indian Journal of Fisheries; Vol. 71 No. 2 (2024)
0970-6011
 
Language eng
 
Relation https://epubs.icar.org.in/index.php/IJF/article/view/127966/54948
 
Rights Copyright (c) 2024 Indian Journal of Fisheries