Please use this identifier to cite or link to this item: http://ir.swu.ac.th/jspui/handle/123456789/14593
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dc.contributor.authorKunanopparat A.
dc.contributor.authorChaivisuthangkura P.
dc.contributor.authorSenapin S.
dc.contributor.authorLongyant S.
dc.contributor.authorRukpratanporn S.
dc.contributor.authorFlegel T.W.
dc.contributor.authorSithigorngul P.
dc.date.accessioned2021-04-05T03:35:51Z-
dc.date.available2021-04-05T03:35:51Z-
dc.date.issued2011
dc.identifier.issn1660934
dc.identifier.other2-s2.0-78650534660
dc.identifier.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-78650534660&doi=10.1016%2fj.jviromet.2010.10.015&partnerID=40&md5=ffe24286974899bb5bddcd9f7325d1d6
dc.identifier.urihttp://ir.swu.ac.th/jspui/handle/123456789/14593-
dc.description.abstractThe 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.
dc.subjectcapsid protein
dc.subjectcapsid protein C
dc.subjectcapsid protein I
dc.subjectcapsid protein N
dc.subjectglutathione transferase
dc.subjectintein
dc.subjectmonoclonal antibody
dc.subjectrecombinant protein
dc.subjectunclassified drug
dc.subjectanimal cell
dc.subjectanimal tissue
dc.subjectantibody specificity
dc.subjectarticle
dc.subjectblood cell
dc.subjectconnective tissue
dc.subjectcontrolled study
dc.subjectDensovirus
dc.subjectgene amplification
dc.subjectgill
dc.subjectheart
dc.subjectheterologous expression
dc.subjectimmunohistochemistry
dc.subjectInfectious myonecrosis virus
dc.subjectlymphoid organ
dc.subjectmouse
dc.subjectmuscle
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectopen reading frame
dc.subjectPolyhedrosis virus
dc.subjectpriority journal
dc.subjectsensitivity analysis
dc.subjectshrimp
dc.subjectTaura syndrome virus
dc.subjectvirus
dc.subjectvirus detection
dc.subjectvirus genome
dc.subjectWestern blotting
dc.subjectWhite spot syndrome virus
dc.subjectYellow head virus
dc.subjectAnimals
dc.subjectAntibodies, Monoclonal
dc.subjectAntibodies, Viral
dc.subjectCapsid Proteins
dc.subjectCloning, Molecular
dc.subjectEscherichia coli
dc.subjectGene Expression
dc.subjectImmunoassay
dc.subjectImmunohistochemistry
dc.subjectMice
dc.subjectPenaeidae
dc.subjectRecombinant Proteins
dc.subjectRNA Viruses
dc.subjectSensitivity and Specificity
dc.subjectVirology
dc.subjectCrangon crangon
dc.subjectDecapoda (Crustacea)
dc.subjectDensovirus
dc.subjectLitopenaeus stylirostris
dc.subjectLitopenaeus vannamei
dc.subjectMiridae
dc.subjectMus
dc.subjectNucleopolyhedrovirus
dc.subjectPenaeidae
dc.subjectPenaeus monodon
dc.subjectShrimp white spot syndrome virus
dc.subjectTaura syndrome virus
dc.subjectYellow head virus
dc.titleDetection of infectious myonecrosis virus using monoclonal antibody specific to N and C fragments of the capsid protein expressed heterologously
dc.typeArticle
dc.rights.holderScopus
dc.identifier.bibliograpycitationJournal of Virological Methods. Vol 171, No.1 (2011), p.141-148
dc.identifier.doi10.1016/j.jviromet.2010.10.015
Appears in Collections:Scopus 1983-2021

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