Please use this identifier to cite or link to this item: https://ir.swu.ac.th/jspui/handle/123456789/14364
ชื่อเรื่อง: Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein
ผู้แต่ง: Wangman P.
Senapin S.
Chaivisuthangkura P.
Longyant S.
Rukpratanporn S.
Sithigorngul P.
Keywords: antibody
biological production
coliform bacterium
crustacean
detection method
disease incidence
enzyme activity
gene expression
immunoassay
medicine
polymerase chain reaction
protein
virus
Crangon crangon
Decapoda (Crustacea)
Escherichia coli
Macrobrachium rosenbergii
Macrobrachium rosenbergii nodavirus
Miridae
Nodaviridae
capsid protein
monoclonal antibody
recombinant protein
virus antibody
animal
animal disease
article
fish disease
genetics
immunology
metabolism
mouse
Nodavirus
Palaemonidae
RNA virus infection
sensitivity and specificity
test strip
virology
Animals
Antibodies, Monoclonal
Antibodies, Viral
Capsid Proteins
Fish Diseases
Mice
Nodaviridae
Palaemonidae
Reagent Strips
Recombinant Proteins
RNA Virus Infections
Sensitivity and Specificity
วันที่เผยแพร่: 2012
บทคัดย่อ: The gene encoding the capsid protein of Macrobrachium rosenbergii nodavirus (MrNV) was cloned into pGEX-6P-1 expression vector and then transformed into the Escherichia coli strain BL21. After induction, capsid protein-glutathione-S- transferase (GST-MrNV; 64 kDa) was produced. The recombinant protein was separated using SDS-PAGE, excised from the gel, electro-eluted and then used for immunization for monoclonal antibody (MAb) production. Four MAbs specific to the capsid protein were selected and could be used to detect natural MrNV infections in M. rosenbergii by dot blotting, Western blotting and immunohistochemistry without cross-reaction with uninfected shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was 10 fmol μl-1 of the GST-MrNV, as determined using dot blotting. However, the sensitivity of the MAb on dot blotting with homogenate from naturally infected M. rosenbergii was approximately 200-fold lower than that of 1-step RT-PCR. Immunohistochemical analysis using these MAbs with infected shrimp tissues demonstrated staining in the muscles, nerve cord, gill, heart, loose connective tissue and inter-tubular tissue of the hepatopancreas. Although the positive reactions occurred in small focal areas, the immunoreactivity was clearly demonstrated. The MAbs targeted different epitopes of the capsid protein and will be used to develop a simple immunoassay strip test for rapid detection of MrNV. © Inter-Research 2012.
URI: https://ir.swu.ac.th/jspui/handle/123456789/14364
https://www.scopus.com/inward/record.uri?eid=2-s2.0-84861668619&doi=10.3354%2fdao02431&partnerID=40&md5=6194606f5f550e7a7b3b1b4c821c35c2
ISSN: 1775103
Appears in Collections:Scopus 1983-2021

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