Detection of lymphocystis disease virus (LCDV) in olive flounder (Paralichthys olivaceus) using efficient extraction and concentration methods

Abstract

Efficiency of extraction and concentration methods for the detection of the major capsid protein gene of lymphocystis disease virus from different tissues of olive flounders (Paralichthys olivaceus) was tested. Tris elution buffer showed a 100 fold higher polymerase chain reaction (PCR) detection limit than TE elution buffer in the virus extraction step from skin tissues. Using the TRPD (Tris elution buffer, polyethylene, and DNA extraction kit) procedure, we confirmed that skin tissues and lymphocystis cells of olive flounders had a detection limit of 10 �6 and 10 �7 PCR-U/µL, meaning that 10 6 and 10 7 fold dilutions of lymphocystis disease virus (LCDV) were PCR positive, respectively.