Detection and titration of white spot syndrome virus using a Blue‐Cell ELISA

Shih, Hsiu-Hui

doi:10.1046/j.1365-2761.2002.00354.x

Cited by 6 publications

(2 citation statements)

References 16 publications

(33 reference statements)

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“…Early detection of WSSV is important and essential for the management of white spot disease (WSD). The various methods for detecting WSSV include histopathology (Takahashi et al 1994;Lightner et al 1996;Mohan et al 1997;Rajendran et al 1999), immunological methods (Nadala & Loh 2000;Poulos et al 2001;Anil, Shankar & Mohan 2002;Liu et al 2002;Shih 2002;You et al 2002;Makesh et al 2006;Sithigorngul et al 2006;Wang, Zhan & Xing 2006), hybridization with gene probes (Chang et al 1996;Durand et al 1996;Nunan & Lightner 1997) and polymerase chain reaction (PCR) (Lo et al 1996b;Takahashi et al 1996;Kim et al 1998;Durand & Lightner 2002;Kono et al 2004). There are very few reports on the development of rapid field-level diagnostics for the sensitive detection of WSSV.…”

Section: Introductionmentioning

confidence: 99%

Development of a monoclonal antibody‐based flow‐through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon

Patil

Shankar

Kumar

et al. 2013

Journal of Fish Diseases

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A flow-through immunoassay (FTA), an improved version of immunodot, was developed using a nitrocellulose membrane baked onto adsorbent pads enclosed in a plastic cassette to detect white spot syndrome virus (WSSV) in shrimp. Sharp purple dots developed with WSSV against the white background of the nitrocellulose membrane. The detection limits of WSSV by the FTA and immunodot were 0.312 and 1.2 μg mL(-1) crude WSSV protein, respectively. The FTA could be completed in 8-10 min compared with 90 min for immunodot. The FTA was 100 times more sensitive than 1-step polymerase chain reaction (PCR) and in between that of the 1- and 2-step PCR protocol recommended by the Office of International Epizootics (OIE). In experimental, orally infected shrimp post-larvae, WSSV was first detected 14, 16 and 18 h post-infection (hpi) by FTA, immunodot and one-step PCR, respectively. The FTA detected WSSV 2 and 4 h earlier than immunodot and one-step PCR, respectively. The FTA was more sensitive (25/27) than one-step PCR (23/27) and immunodot (23/27) for the detection of WSSV from white spot disease outbreak ponds. The reagent components of the FTA were stable giving expected results for 6 m at 4-8 °C. The FTA is available as a rapid test kit called 'RapiDot' for the early detection of WSSV under field conditions.

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Section: Introductionmentioning

confidence: 99%

Development of a monoclonal antibody‐based flow‐through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon

Patil

Shankar

Kumar

et al. 2013

Journal of Fish Diseases

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“…Unfortunately, the lack of immortal crustacean cell lines has been hampering the in vitro determination of WSSV infectious titers. The use of primary crustacean cell culture systems for titration of WSSV has been investigated, but these systems have not yet reached a sufficient level of reproducibility and standardization [174][175][176][177]. Consequently, standardized methods were published to determine the shrimp infectious dose 50% endpoint (SID 50 ) and the lethal infectious dose 50% endpoint (LD 50 ) during controlled in vivo titration experiments [51,53,105].…”

Section: Virulence According To the Strainmentioning

confidence: 99%

Experimental Infection Models and Their Usefulness for White Spot Syndrome Virus (WSSV) Research in Shrimp

Cox,

De Swaef,

Corteel

et al. 2024

Viruses

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White spot syndrome virus (WSSV) is marked as one of the most economically devastating pathogens in shrimp aquaculture worldwide. Infection of cultured shrimp can lead to mass mortality (up to 100%). Although progress has been made, our understanding of WSSV’s infection process and the virus–host–environment interaction is far from complete. This in turn hinders the development of effective mitigation strategies against WSSV. Infection models occupy a crucial first step in the research flow that tries to elucidate the infectious disease process to develop new antiviral treatments. Moreover, since the establishment of continuous shrimp cell lines is a work in progress, the development and use of standardized in vivo infection models that reflect the host–pathogen interaction in shrimp is a necessity. This review critically examines key aspects of in vivo WSSV infection model development that are often overlooked, such as standardization, (post)larval quality, inoculum type and choice of inoculation procedure, housing conditions, and shrimp welfare considerations. Furthermore, the usefulness of experimental infection models for different lines of WSSV research will be discussed with the aim to aid researchers when choosing a suitable model for their research needs.

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Bacterial Lipid Modification of ICP11 and a New ELISA System Applicable for WSSV Infection Detection

Murugan

Sankaran

2018

Mar Biotechnol

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In ELISA, a popular analytical diagnostic tool, the stable non-covalent immobilization (coating) of hydrophilic proteins/peptides on to hydrophobic polystyrene surface has remained a major common challenge. Recombinant bacterial lipid modification of proteins in Escherichia coli system has been shown in this study to solve this problem owing to the hydrophobic anchorage provided by three fatty acyl groups in N-acyl-S-diacylglyceryl Cys at the N-terminus. Exploiting this first post-translational protein engineering, the most abundantly expressed white spot syndrome viral protein ICP11 was lipid-modified and tested as a new target in a new ELISA method useful to shrimp farming. The lipid served as a potent adjuvant to enhance the titer (16 times) of higher affinity antibodies where amino terminal lipoamino acid N-acyl-S-diacylglyceryl cysteine of bacterial lipoproteins induce inflammatory responses through TLR and stimulate humoral immune responses without additional adjuvant and also aided in the immobilization of even a few nanograms of ICP11. Competition between the immobilized and the free antigen from the sample provided a sensitive measure of antigen in the infected shrimp tissues. The detection limit for ICP11 protein using competitive ELISA was 250 pg and the linear range of the assay was 15-240 ng.

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Detection and titration of white spot syndrome virus using a Blue‐Cell ELISA

Cited by 6 publications

References 16 publications

Development of a monoclonal antibody‐based flow‐through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon

Development of a monoclonal antibody‐based flow‐through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon

Experimental Infection Models and Their Usefulness for White Spot Syndrome Virus (WSSV) Research in Shrimp

Bacterial Lipid Modification of ICP11 and a New ELISA System Applicable for WSSV Infection Detection

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