Publication: Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11
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Issued Date
2013
Resource Type
File Type
application/pdf
ISSN
3048608
Other identifier(s)
2-s2.0-84876719031
Rights Holder(s)
Scopus
Bibliographic Citation
Archives of Virology. Vol 158, No.5 (2013), p.967-979
Suggested Citation
Siriwattanarat R., Longyant S., Chaivisuthangkura P., Wangman P., Vaniksampanna A., Sithigorngul P. Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11. Archives of Virology. Vol 158, No.5 (2013), p.967-979. doi:10.1007/s00705-012-1569-3 Retrieved from: https://hdl.handle.net/20.500.14740/6665
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. © 2012 Springer-Verlag Wien.