Molecular methods for the diagnosis of Marteilia refringens

A nested polymerase chain reaction (PCR) has been developed for the diagnosis of Marteilia refringens in mussels (Mytilus galloprovincialis) from Galicia (NW Spain). Specific primers were used in two consecutive amplification reactions, that were able to detect 15 fg of Marteilia DNA. The nested PCR yielded a 550 bp fragment that was used to generate a non-radioactive probe, specific for Marteilia, as determined by in situ hybridization. This probe was also used in a dot-blot developed for the diagnosis of the parasite. This method alone was much less sensitive than the nested PCR, however, its use after the nested PCR significantly increased the sensitivity of the PCR. When comparing these molecular diagnosis tools with traditional diagnosis methods (smears and histology), we determined that the nested PCR could detect Marteilia in 65% of the mussels screened, while only 20-25% of the mussels were identified as infected by traditional methods.TR: MB0200756Source type: Electronic(1