Development and application of monoclonal antibodies for the detection of white spot syndrome virus of penaeid shrimp

Abstract

Monoclonal antibodies (MAbs) were produced against white spot syndrome virus (WSSV) of penaeid shrimp. The virus isolate used for immunization was obtained from China in 1994 and was passaged in Penaeus vannamei. The 4 hybridomas selected for characterization all produced MAbs that reacted with the 28 kD structural protein by Western blot analysis. The MAbs tested in dot-immunoblot assays were capable of detecting the virus in hemolymph samples collected from moribund shrimp during an experimentally induced WSSV infection. Two of the MAbs were chosen for development of serological detection methods for WSSV. The 2 MAbs detected WSSV infections in fresh tissue impression smears using a fluorescent antibody for final detection. A rapid immunohistochemical method using the MAbs on Davidson's fixed tissue sections identified WSSV-infected cells and tissues in a pattern similar to that seen with digoxigenin-labeled WSSV-specific gene probes. A whole mount assay of pieces of fixed tissue without paraffin embedding and sectioning was also successfully used for detecting the virus. None of the MAbs reacted with hemolymph from specific pathogen-free shrimp or from shrimp infected with infectious hypodermal and hematopoietic necrosis virus, yellow head virus or Taura syndrome virus. In Western blot analysis, the 2 MAbs did not detect any serological differences among WSSV isolates from China, Thailand, India, Texas, South Carolina or Panama. Additionally, the MAbs did not detect a serological difference between WSSV isolated from penaeid shrimp and WSSV isolated from freshwater crayfish. KEY WORDS: White spot syndrome virus · Monoclonal antibodies · ImmunodetectionResale or republication not permitted without written consent of the publisher

Construction and characterization of a novel recombinant single-chain variable fragment antibody against White Spot Syndrome Virus from shrimp

“…Many investigators have studied this virus to try to control the massive disease outbreak. Diagnostic tools, such as gene probes (Duarand et al, 1996), polymerase chain reaction (PCR) (Lo et al, 1996) and immunological methods (Poulos et al, 2001;Nadala and Loh, 2000), have been developed to control the spread of WSSV from parent shrimps and imported shrimps.…”

Production of recombinant enveloped structural proteins from the Chinese WSSV isolate

“…The VP19 and VP28 genes were selected for cloning in pUCm-T vector and sub cloned in pET-30a (+) vector and the recombinant proteins have been produced, as both these genes code for envelope protein (15,19) and they showed few base pair changes. This therefore suggests that VP19 protein may be involved in pathogenesis, and further investigations need to be carried out to substantiate this view.…”

“…Various kinds of rapid diagnostic tools, such as gene probes (19), PCR (18) and immunological methods (20)(21)(22) have been developed to detect the spread of WSSV. The immunological methods are not so accurate and sensitive as compared to PCR but are convenient for the field level detection of WSSV.…”

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“…The same group described production of MAb to ScienceAsia ScienceAsia ScienceAsia ScienceAsia 32 (2006) 32 (2006) 32 (2006) 32 (2006) 32 (2006) the 28 kDa envelope protein of white spot syndrome virus (WSSV) and its use for immunodiagnosis. 24 Sithigorngul et al 20 reported the production of monoclonal antibodies specific to 22, 64, and 135 kDa proteins of yellow-head virus (YHV) and they have been used for viral detection 20 and typing 25 . We can now add MBV to the list of shrimp viruses for which MAb detection reagents are available.…”