Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp 11

Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific... The icp 11 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp 11

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Publisher
Springer Vienna
Copyright
Copyright © 2013 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-012-1569-3
Publisher site
See Article on Publisher Site

References

  • Detection of major penaeid shrimp viruses in Asia, a historical perspective with emphasis on Thailand
    Flegel, TW
  • Qualitative and quantitative studies on the relative virus load of tails and heads of shrimp acutely infected with WSSV
    Durand, SV; Redman, RM; Mohney, LL; Tang-Nelson, K; Bonami, JR; Lightner, DV
  • Detection of white spot syndrome virus in shrimp by loop-mediated isothermal amplification
    Kono, T; Savan, R; Sakai, M; Itami, T
  • Real-time quantitative loop-mediated isothermal amplification as a simple method for detecting white spot syndrome virus
    Mekata, T; Sudhakaran, R; Kono, T; Supamattaya, K; Linh, NT; Sakai, M; Itami, T
  • Rapid and sensitive detection of white spot syndrome virus by loop-mediated isothermal amplification combined with a lateral flow dipstick
    Jaroenram, W; Kiatpathomchai, W; Flegel, TW

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