Please use this identifier to cite or link to this item: http://ir.swu.ac.th/jspui/handle/123456789/14593
Title: Detection of infectious myonecrosis virus using monoclonal antibody specific to N and C fragments of the capsid protein expressed heterologously
Authors: Kunanopparat A.
Chaivisuthangkura P.
Senapin S.
Longyant S.
Rukpratanporn S.
Flegel T.W.
Sithigorngul P.
Keywords: capsid protein
capsid protein C
capsid protein I
capsid protein N
glutathione transferase
intein
monoclonal antibody
recombinant protein
unclassified drug
animal cell
animal tissue
antibody specificity
article
blood cell
connective tissue
controlled study
Densovirus
gene amplification
gill
heart
heterologous expression
immunohistochemistry
Infectious myonecrosis virus
lymphoid organ
mouse
muscle
nonhuman
nucleotide sequence
open reading frame
Polyhedrosis virus
priority journal
sensitivity analysis
shrimp
Taura syndrome virus
virus
virus detection
virus genome
Western blotting
White spot syndrome virus
Yellow head virus
Animals
Antibodies, Monoclonal
Antibodies, Viral
Capsid Proteins
Cloning, Molecular
Escherichia coli
Gene Expression
Immunoassay
Immunohistochemistry
Mice
Penaeidae
Recombinant Proteins
RNA Viruses
Sensitivity and Specificity
Virology
Crangon crangon
Decapoda (Crustacea)
Densovirus
Litopenaeus stylirostris
Litopenaeus vannamei
Miridae
Mus
Nucleopolyhedrovirus
Penaeidae
Penaeus monodon
Shrimp white spot syndrome virus
Taura syndrome virus
Yellow head virus
Issue Date: 2011
Abstract: The gene encoding the capsid protein in ORF1 of the genome of infectious myonecrosis virus (IMNV) (GenBank AY570982) was amplified into three parts named CP-N (nucleotides 2248-3045), CP-I (nucleotides 3046-3954) and CP-C (nucleotides 3955-4953). The CP-N fragment was inserted into expression vector pTYB1 while CP-I and CP-C were each inserted into expression vector pGEX-6P-1 for transformation of BL21 E. coli strain. After induction, intein-CP-N (84 kDa), glutathione-S-transferase (GST)-CP-I (60 kDa) and GST-CP-C (62 kDa) fusion proteins were produced. They were separated by SDS-PAGE and electroeluted before immunization of Swiss mice for monoclonal antibody (MAb) production. Two MAbs specific to CP-N and one MAb specific to CP-C were selected for use for detection of natural IMNV infections in Penaeus vannamei by dot blotting, Western blotting and immunohistochemistry. There was no cross-reaction with shrimp tissues or common shrimp viruses including white spot syndrome virus (WSSV), yellow head virus (YHV), Taura syndrome virus (TSV), Penaeus monodon nucleopolyhedrovirus (PemoNPV), Penaeus stylirostris densovirus (PstDNV) and Penaeus monodon densovirus (PmDNV). The detection sensitivities of the MAbs were approximately 6 fmol/spot of purified recombinant intein-CP-N protein and 8 fmol/spot of GST-CP-C as determined by dot blotting. A combination of all three MAbs resulted in a twofold increase in sensitivity over use of any single MAb. However, this sensitivity was approximately 10 times lower than that of one-step RT-PCR using the same sample. Immunohistochemical analysis using MAbs specific to CP-N and CP-C in IMNV-infected shrimp revealed intense staining patterns in muscles, the lymphoid organ, gills, the heart, hemocytes and connective tissue. © 2010 Elsevier B.V.
URI: https://www.scopus.com/inward/record.uri?eid=2-s2.0-78650534660&doi=10.1016%2fj.jviromet.2010.10.015&partnerID=40&md5=ffe24286974899bb5bddcd9f7325d1d6
http://ir.swu.ac.th/jspui/handle/123456789/14593
ISSN: 1660934
Appears in Collections:Scopus 1983-2021

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