Publication:
Detection of infectious myonecrosis virus using monoclonal antibody specific to N and C fragments of the capsid protein expressed heterologously

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Issued Date

2011

Resource Type

Article

File Type

application/pdf

ISSN

1660934

DOI

10.1016/j.jviromet.2010.10.015

Other identifier(s)

2-s2.0-78650534660

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of Virological Methods. Vol 171, No.1 (2011), p.141-148

Suggested Citation

Kunanopparat A., Chaivisuthangkura P., Senapin S., Longyant S., Rukpratanporn S., Flegel T.W., Sithigorngul P. Detection of infectious myonecrosis virus using monoclonal antibody specific to N and C fragments of the capsid protein expressed heterologously. Journal of Virological Methods. Vol 171, No.1 (2011), p.141-148. doi:10.1016/j.jviromet.2010.10.015 Retrieved from: https://hdl.handle.net/20.500.14740/7426

Title

Detection of infectious myonecrosis virus using monoclonal antibody specific to N and C fragments of the capsid protein expressed heterologously

Author(s)

Kunanopparat A.
 Chaivisuthangkura P.
 Senapin S.
 Longyant S.
 Rukpratanporn S.
 Flegel T.W.
 Sithigorngul P.

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.

Subject(s)

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

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URI

https://hdl.handle.net/20.500.14740/7426

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Scopus: Year 1983-2021

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