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Title: Simultaneous and rapid detection of white spot syndrome virus and yellow head virus infection in shrimp with a dual immunochromatographic strip test
Authors: Sithigorngul P.
Rukpratanporn S.
Chaivisuthangkura P.
Sridulyakul P.
Longyant S.
Keywords: monoclonal antibody
virus protein
animal experiment
controlled study
diagnostic test accuracy study
immunochromatographic strip test
intermethod comparison
priority journal
reverse transcription polymerase chain reaction
RNA virus
RNA virus infection
sensitivity and specificity
test strip
virus detection
White spot syndrome virus
Yellow head virus
Yellow head virus infection
Antibodies, Monoclonal
Antibodies, Viral
Sensitivity and Specificity
White spot syndrome virus 1
Capra hircus
Decapoda (Crustacea)
Litopenaeus vannamei
Shrimp white spot syndrome virus
Yellow head virus
Issue Date: 2011
Abstract: A strip test for the dual detection of white spot syndrome virus (WSSV) and yellow head virus (YHV) was developed using monoclonal antibodies (MAbs) specific to the WSSV major envelope protein VP28 (W1 and W30) and the YHV nucleocapsid protein p20 (Y19 and Y21). The MAbs W30 and Y19 were conjugated with colloidal gold and sprayed onto a glass fiber pad that was placed adjacent to a sample chamber. The MAbs W1 and Y21 and the goat anti-mouse immunoglobulin G (GAM) antibody were sprayed onto a nitrocellulose membrane in strips at positions designated W, Y and C, respectively. These test strips were placed in plastic cases and stored desiccated in a plastic bag. The test strips were assessed for their ability to detect WSSV and YHV simultaneously using pleopods sampled from shrimp. A pleopod homogenate in application buffer 100μl was applied to the sample chamber to flow through the nitrocellulose membrane strip, and antibody-protein complexes could be observed within 15. min. In sample from shrimp infected with WSSV and/or YHV, viral protein bound to the colloidal gold-conjugated MAbs. These complexes were captured by the MAbs at the W and/or Y test lines, resulting in the appearance of reddish-purple coloured bands. Any unbound colloidal gold-conjugated MAbs migrated pass the W and Y lines would be captured by the GAM antibody, forming a band at position C. When samples not containing WSSV and YHV proteins or containing viral proteins at below the detection limit of the test, only the band at position C was observed. The sensitivity of the test was comparable to dot blot tests using single MAbs, and ~500-fold less sensitive than a 1-step PCR test for WSSV and 1000-fold less sensitive than an RT-PCR test for YHV. Despite this lower sensitivity, the dual strip test has advantages in speed and simplicity in not requiring sophisticated equipment or specialized skills. The ability to co-detect WSSV and YHV provides simultaneously cost savings. © 2011 Elsevier B.V.
ISSN: 1660934
Appears in Collections:SCOPUS 1983-2021

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