ABSTRACT
A simple reverse passive latex agglutination (RPLA) method for detecting white spot syndrome virus (WSSV) in the hemolymph of infected Kuruma shrimp (Penaeus japonicus) was developed. It was confirmed that WSSV could be detected from the shrimp hemolymph when the latex particles blocked with a casein protein were used as detection reagent. It became clear from the result of the infection trial that viruses are detectable by RPLA before the appearance of overt symptoms of this disease. In addition, an amplification product of 982 bp (s) derived from WSSV by PCR was detected in all the samples in which WSSV was detected by RPLA. This newly developed RPLA assay can examine many samples in a simple manner since hemolymph can be extracted more easily than any other organs. This assay can be used conveniently for virus detection in the culture pond of shrimps or in the field.
Subject(s)
Hemolymph/virology , Latex Fixation Tests/methods , Penaeidae/virology , White spot syndrome virus 1/isolation & purification , Animals , Chromatography, Gel , Chromatography, Ion Exchange , Female , Male , Polymerase Chain ReactionABSTRACT
A reversed passive latex agglutination (RPLA) assay was developed for detecting the white spot syndrome virus (WSSV), which was formally named as penaeid rod-shaped DNA virus (PRDV) in Japan, from stomach tissue homogenate of the kuruma shrimp (Penaeus japonicus). Using high-density latex particles and specific polyclonal antibody, WSSV was detectable after 4h incubation. The hemolymph, the stomach, and the gills were extracted from a shrimp that had been infected experimentally with WSSV, the virus contained in each sample was tested by the PRLA and PCR assay. It was possible to detect the WSSV only from stomach tissue homogenates by the RPLA assay. And there was an agreement between RPLA and PCR assays for WSSV detection. Considering that the RPLA assay does not require biochemical expertise and latex reagents and all apparatus can be provided as a kit, this assay can be used for virus detection in the culture pond of shrimps or in the field as a convenient method.
