DAO 99:197-205 (2012)  -  DOI: https://doi.org/10.3354/dao02482

Monoclonal antibodies against extra small virus show that it co-localizes with Macrobrachium rosenbergii nodavirus

Siwaporn Longyant1, Saengchan Senapin2,3, Sirijantra Sanont1, Pradit Wangman1, Parin Chaivisuthangkura1, Sombat Rukpratanporn4, Paisarn Sithigorngul1,*

1Department of Biology, Srinakharinwirot University, Bangkok 10110, Thailand
2CENTEX Shrimp, Mahidol University, Bangkok 10400, Thailand
3National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathumthani 12120, Thailand
4Center of Excellence for Marine Biotechnology at Chulalongkorn University, National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok 10330, Thailand
*‑Corresponding author. Email:

ABSTRACT: The capsid protein (CP) gene of extra small virus (XSV) expressed in Escherichia coli as a 42 kDa glutathione S-transferase (GST)-fusion protein (GST-XCP) or a 20 kDa His6-fusion protein (His6-XCP) were purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), combined, and used to immunize Swiss mice to produce monoclonal antibodies (MAbs). Using dot blot, Western blot, and immunohistochemistry (IHC) methods, 4 MAbs specific to the XSV CP detected XSV in the freshwater prawn Macrobrachium rosenbergii without cross-reaction to host proteins or to proteins of Macrobrachium rosenbergii nodavirus (MrNV) or 5 of the most pathogenic viruses of penaeid shrimp. In dot blots, the combined MAbs could detect down to ~10 to 20 fmol ┬Ál−1 of purified GST-XCP protein, which was somewhat more sensitive compared to any single MAb. Used in conjunction with an MrNV-specific MAb, white tail disease (WTD) was diagnosed more effectively. However, the sensitivity at which the combined 4 MAbs detected XSV CP was 1000-fold lower than XSV RNA detected by RT-PCR. IHC analysis of M. rosenbergii tissue sections using the MAbs showed XSV infection to co-localize at variable loads with MrNV infection in heart and muscle cells as well as cells of connective tissues in the hepatopancreas. Since XSV histopathology remained prominent in tissues of some prawns in which MAb reactivity for MrNV was low compared to MAb reactivity for XSV, XSV might play some role in WTD severity.


KEY WORDS: Immunohistochemistry · Macrobrachium rosenbergii nodavirus · MrNV · Monoclonal antibody · Capsid protein · Western blot · Extra small virus · XSV


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Cite this article as: Longyant S, Senapin S, Sanont S, Wangman P, Chaivisuthangkura P, Rukpratanporn S, Sithigorngul P (2012) Monoclonal antibodies against extra small virus show that it co-localizes with Macrobrachium rosenbergii nodavirus. Dis Aquat Org 99:197-205. https://doi.org/10.3354/dao02482

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