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Title: Rapid and sensitive detection of Penaeus monodon nucleopolyhedrovirus by loop-mediated isothermal amplification
Authors: Chaivisuthangkura P.
Srisuk C.
Rukpratanporn S.
Longyant S.
Sridulyakul P.
Sithigorngul P.
Keywords: genomic DNA
analytical equipment
DNA extraction
Infectious hematopoietic necrosis virus
loop mediated isothermal amplification
nucleic acid amplification
nucleotide sequence
Penaeus monodon nucleopolyhedrovirus
Penaeus stylirostris densovirus
Polyhedrosis virus
polymerase chain reaction
priority journal
sensitivity analysis
Taura syndrome virus
virus detection
virus gene
virus infection
White spot syndrome virus
yellow head virus
DNA Primers
Nucleic Acid Amplification Techniques
Polymerase Chain Reaction
Sensitivity and Specificity
Species Specificity
Time Factors
Viral Structural Proteins
Decapoda (Crustacea)
Hepatopancreatic parvovirus of penaeid shrimp
Infectious hypodermal and hematopoietic necrosis virus
Litopenaeus stylirostris
Monodon baculovirus
Penaeus monodon
Shrimp white spot syndrome virus
Taura syndrome virus
Yellow head virus
Issue Date: 2009
Abstract: Loop-mediated isothermal amplification (LAMP) is a novel, sensitive and rapid method for amplification of nucleic acids under isothermal conditions. In this report, a LAMP method was developed for detection of Penaeus monodon nucleopolyhedrovirus (PemoNPV), known previously as monodon baculovirus (MBV), using a set of six primers designed to specifically recognize the PemoNPV polyhedrin gene. The optimized time and temperature conditions for the LAMP assay were 60 min at 63 °C. The sensitivity of LAMP for PemoNPV detection was approximately 50 viral copies ng-1 genomic DNA (equivalent to 150 viral copies per reaction). Using a DNA template extracted from PemoNPV-infected shrimp by a viral nucleic acid kit, the detection limit of LAMP was 0.7 fg while that of nested PCR was 70 fg; therefore, the LAMP assay was 100 times more sensitive than nested PCR. The LAMP method did not amplify a product using nucleic acid extracted from shrimp infected with other viruses including yellow head virus (YHV), Taura syndrome virus (TSV), white spot syndrome virus (WSSV), Penaeus stylirostris densovirus (PstDNV) known previously as infectious hypodermal and hematopoietic necrosis virus (IHHNV), and Penaeus monodon densovirus (PmDNV) known previously as hepatopancreatic parvovirus (HPV). © 2009 Elsevier B.V. All rights reserved.
ISSN: 1660934
Appears in Collections:Scopus 1983-2021

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