T. Renault et al., Development of a PCR procedure for the detection of a herpes-like virus infecting oysters in France, J VIROL MET, 88(1), 2000, pp. 41-50
A PCR-based procedure for detecting a herpes-like virus that infects the Ja
panese oyster, Crassostrea gigas, in France was developed. Two primers were
designed to provide specific amplification products ranging in size from 9
17 to 1001 bp when carried out on oyster herpes-like virus DNA. No amplific
ation was observed of oyster genomic DNA nor of the DNA from vertebrate her
pesviruses. Crude samples were prepared and submitted to nested PCR, allowi
ng amplification of DNA fragments of the expected size when carried out on
infected larval and spat samples. The procedure used to prepare the sample
for PCR was found to be critical because of the presence of unidentified su
bstances in oyster tissues that inhibit the PCR reaction. A rapid and conve
nient sample preparation using ground tissues allowed a sensitive detection
of the herpes-like virus infected oysters. The ability of the defined PCR
protocol to diagnose herpes-like virus infections in oysters was compared t
o the transmission electron microscopy technique using 15 C. gigas larval b
atches with or without mortalities. PCR amplification is as sensitive a dia
gnostic assay for herpes-like virus as transmission electron microscopy. Ho
wever, the nested PCR protocol is more convenient and less time consuming.
The relationship between reported mortalities among C. gigas oyster spat an
d herpes-like virus DNA detection by PCR was also investigated. Statistical
analysis showed that virus detection and mortalities are correlated. This
observation highlights the importance of studying the causative role of her
pes-like virus in oyster spat mortalities. (C) 2000-Elsevier Science B.V. A
ll rights reserved.