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Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11

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Abstract

The icp11 gene encoding the highly abundant DNA mimic protein of white spot syndrome virus (WSSV) was cloned into the pTYB1 and pGEX-6P-1 expression vectors and introduced into E. coli by transformation. After induction, C-terminally intein-tagged ICP11 (ICP11-intein) and N-terminally glutathione-S-transferase (GST)-tagged ICP11 (GST-ICP11) proteins with molecular masses of 64 and 35 kDa were obtained. These proteins were purified by SDS-PAGE and used for immunization of Swiss mice for monoclonal antibody (MAb) production. Two MAbs specific for ICP11 were selected; these MAbs can be used to detect natural WSSV infection in Penaeus vannamei by dot blotting, western blotting or immunohistochemistry without cross-reaction with other shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was approximately 0.7 fmole/spot of GST-ICP11 as determined by dot blotting. These MAbs showed stronger immunoreactivity than other MAbs from previous studies that are specific for VP28 and VP19. A combination of MAbs specific for ICP11, VP28 and VP19 increased the detection sensitivity of WSSV during early infection to a sensitivity 250 times lower than that of one-step PCR. Therefore, the MAbs specific for ICP11 could be used to confirm and enhance the detection sensitivity for WSSV infection in shrimp using various types of antibody-based assays.

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Acknowledgments

This work was financially supported by the Strategic Wisdom and Research Institute of Srinakharinwirot University and the Higher Commission of Education, Ministry of Education, by a graduate scholarship to Mrs. Ruthairat Siriwattanarat.

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Authors and Affiliations

  1. Department of Biology, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Ruthairat Siriwattanarat, Siwaporn Longyant, Parin Chaivisuthangkura, Pradit Wangman, Akapon Vaniksampanna & Paisarn Sithigorngul

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  1. Ruthairat Siriwattanarat
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Correspondence to Paisarn Sithigorngul.

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Siriwattanarat, R., Longyant, S., Chaivisuthangkura, P. et al. Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11. Arch Virol 158, 967–979 (2013). https://doi.org/10.1007/s00705-012-1569-3

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  • DOI: https://doi.org/10.1007/s00705-012-1569-3

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