Generation of monoclonal antibodies specific to hepatopancreatic parvovirus (HPV) from Penaeus monodon

Rukpratanporn, Sombat; Sukhumsirichart, Wasana; Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Weerawan; Menasveta, Piamsak; Sithigorngul, Paisarn

doi:10.3354/dao065085

Cited by 26 publications

(12 citation statements)

References 16 publications

(20 reference statements)

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“…After washing with distilled water for 5 min, the sections were re-fixed with 10% formalin for 10 min, and then washed with PBS, pH 7.2, 3 times for 15 min each. After blocking with P 1 + (10% calf serum in PBS, pH 7.2) for 30 min, they were incubated with MAb specific to PstDNV (Bui 2007) or PmDNV (Rukpratanporn et al 2005) for 5 h at 37°C or overnight at 4°C. The MAb was removed by washing the sections in PBS, pH 7.2, 3 times for 10 min each.…”

Section: Methodsmentioning

confidence: 99%

“…Further, the Taiwanese report does not include TEM or enzyme digestion results. Thus, comparison with our work must await further clarification.In addition to in situ hybridization tests, we carried out immunohistochemical tests using antibodies prepared specifically against heterologously expressed proteins of PmDNV (Rukpratanporn et al 2005) and PstDNV (Bui 2007). The results of these tests were also negative, although positive with respective control Based on the target tissue, on the TEM results and on histopathology, the virus described appears to most closely resemble hepanviruses characterized from penaeid shrimp.…”

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confidence: 99%

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Parvo-like virus in the hepatopancreas of freshwater prawns Macrobrachium rosenbergii cultivated in Thailand

Gangnonngiw¹,

Kiatpathomchai²,

Sriurairatana³

et al. 2009

Dis. Aquat. Org.

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A survey of cultivated giant freshwater prawns Macrobrachium rosenbergii from Thailand revealed the presence of unusual spherical to ovoid inclusions in nuclei of hepatopancreas tubule epithelial cells. These began as small eosinophilic inclusions that became more basophilic as they increased in size. They were present in both R-cells and E-cells but were largest and deeply basophilic only in the E-cells. Confocal laser microscopy revealed that stained nucleic acid fluorescence from the inclusions was lost by treatment with DNase I specific for double-and single-stranded DNA and also lost or reduced by treatment with mungbean nuclease specific for single-stranded nucleic acids. Transmission electron microscopy (TEM) revealed that the inclusions contained tightly packed, unenveloped, viral-like particles of approximately 25 to 30 nm diameter, resembling those produced by shrimp parvoviruses. However, PCR, in situ hybridization and immunohistochemical tests for shrimp parvoviruses previously reported from Thailand were all negative. These results suggested that the inclusions contained a parvo-like virus, not previously reported from M. rosenbergii in Thailand. KEY WORDS: Macrobrachium rosenbergii · Hepatopancreas · Parvo-like virus · Histopathology · Electron microscopy Resale or republication not permitted without written consent of the publisherDis Aquat Org 85: [167][168][169][170][171][172][173] 2009 natural infections of M. rosenbergii is similar to the one known as hepatopancreatic parvovirus (HPV) in penaeid shrimp (Anderson et al. 1990, Lightner et al. 1994) based on histopathology (Bonami et al. 1995, Lightner 1996. A recent proposal to the International Committee on Taxonomy of Viruses (Tijssen 2008) recommends that these penaeid shrimp parvoviruses be included in a new genus, Hepanvirus, in the sub-family Densovirinae of the family Parvoviridae as Penaeus monodon densovirus (PmDNV), Penaeus merguiensis densovirus (PmergDNV) and Penaeus chinensis densovirus (PchinDNV). These differ in genome size and organization and target tissue from another penaeid shrimp parvovirus also included in the sub-family Densovirinae, but in the genus Brevidensovirus, as Penaeus stylirostris densovirus (PstDNV) (Tattersall et al. 2005). PstDNV was formerly called infectious hypodermal and hematopoeitic necrosis virus (IHHNV) (Lightner 1996). The newly recommended names (Tattersall et al. 2005, Tijssen 2008) will be used in this manuscript, as will the ICTV method of naming shrimp viruses with non-italicized host species names and abbreviations (Fauquet et al. 2005).For successful specific pathogen free (SPF) stock development, diagnostic methods would be required for all of the pathogens included in any specific list of pathogens for M. rosenbergii. For this species, highly sensitive RT-PCR methods are available for MrNV and XSV (Romestand & Bonami 2003, Sri Widada et al. 2003, Yoganandhan et al. 2005, Pillai et al. 2006. However, for other viral pathogens, such as the parvo-like virus described above, no molec...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Parvo-like virus in the hepatopancreas of freshwater prawns Macrobrachium rosenbergii cultivated in Thailand

Gangnonngiw¹,

Kiatpathomchai²,

Sriurairatana³

et al. 2009

Dis. Aquat. Org.

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“…Shrimp samples infected with Penaeus monodon densovirus (PmDNV; also called hepatopancreatic parvovirus), Penaeus monodon nucleopolyhedrovirus (PemoNPV; also called monodon baculo virus), Taura syndrome virus (TSV), white spot syn drome virus (WSSV), yellow head virus (YHV) and infectious myonecrosis virus (IMNV) were processed for paraffin sectioning and immunohistochemistry using MAbs specific to MrNV. The results were compared to those from MAbs specific to PmDNV (Rukpratanporn et al 2005), TSV (Longyant et al 2008), WSSV (Chaivisuthangkura et al 2004), YHV (Sithigorngul et al 2002), PemoNPV (Boonsanongchokying et al 2006), Penaeus stylirostris densovirus (PstDNV; Sithigorngul et al 2009) and IMNV (Kunanopparat et al 2011). …”

Section: Immunohistochemistrymentioning

confidence: 99%

Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein

Wangman¹,

Senapin²,

Chaivisuthangkura³

et al. 2012

Dis. Aquat. Org.

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The gene encoding the capsid protein of Macrobrachium rosenbergii nodavirus (Mr NV) 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-Mr NV; 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 Mr NV 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-Mr NV, 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 Mr NV. KEY WORDS: Capsid protein · Extra small virus · XSV · Immunohistochemistry · Macrobrachium rosenbergii nodavirus · Mr NV · Monoclonal antibody · Western blot Resale or republication not permitted without written consent of the publisherDis Aquat Org 98: [121][122][123][124][125][126][127][128][129][130][131] 2012 Genome-based detection methods with high specificity and high sensitivity for detection of MrNV include dot blot hybridization, in situ hybridization (Sri Widada et al. 2003) and 1-step RT-PCR in the form of single tests for MrNV or XSV (Sri Widada et al. 2003, Sahul Hameed et al. 2004 or multiplex tests for both viruses (Yoganandhan et al. 2005, Tripathy et al. 2006. A 2-step real-time RT-PCR method (Zhang et al. 2006) and a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method (Pillai et al. 2006, Puthawibool et al. 2010) have also been described. Although a thermal cycler is not required for RT-LAMP, the technique is still expensive and fairly complicated, and a laboratory is required. Therefore, these molecularbased techniques are not useful for pond-side detection by shrimp farmers, who require simpler and quicker disease monitoring and disease-outbreak confirmation methods that are easy to perform with high specificity and optimal sensitivity. These objectives may be achieved using immunoassay methods. Use of a polyclonal antibody (PAb) (Romestand & Bonami 2003) and monoclonal antibodies (MAbs) (Qian et al. 2006) to detect purified MrNV using sandwich ELISA and Western blotting (Sahul Hameed et al. 2011) have been described. Alth...

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“…ORF3 encodes a capsid protein of approximately 92 kDa, which may later be cleaved after the first or second arginine residue to produce a 57 kDa or 54 kDa structural protein, respectively [82] . MAbs raised against purified PmDNV isolated from Thailand have been produced, four of which react with a 54 kDa protein by Western blotting and to intranuclear inclusion bodies in tubule epithelial cells in PmDNVinfected tissue by immunohistochemistry [83] . The recombinant capsid proteins of PmDNV have also been used to generate MAbs, and five MAbs have been obtained.…”

Section: Pmdnvmentioning

confidence: 99%

Immunological-based assays for specific detection of shrimp viruses

Chaivisuthangkura

Longyant

Sithigorngul

2014

WJV

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Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei , and the black tiger shrimp, Penaeus (Penaeus) monodon . Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In

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Generation of monoclonal antibodies specific to hepatopancreatic parvovirus (HPV) from Penaeus monodon

Cited by 26 publications

References 16 publications

Parvo-like virus in the hepatopancreas of freshwater prawns Macrobrachium rosenbergii cultivated in Thailand

Parvo-like virus in the hepatopancreas of freshwater prawns Macrobrachium rosenbergii cultivated in Thailand

Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein

Immunological-based assays for specific detection of shrimp viruses

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