APPLICATION OF A PCR-BASED TECHNIQUE FOR DETECTING AND DISCRIMINATING BETWEEN MYCOPLASMA SPECIES FROM CALIFORNIA SEA LIONS (ZALOPHUS CALIFORNIANUS)

摘要

Bacterial infections are an important cause of morbidity and mortality in marine mammals. The accurate identification of a suspected bacterial pathogen relies on culture followed by time-consuming and subjective phenotypic tests. In human medicine molecular diagnosis is playing an increasingly important role in the rapid detection and identification of pathogenic organisms in clinical samples. The genetic variation of ribosomal genes, which are universally present in bacteria, offers an alternative to culturing for the detection and identification of these organisms. The 16S ribosomal gene is an excellent candidate for molecular bacterial detection since it contains conserved sequence regions ideal for primer targeting as well as regions of variability useful for species identification. We have recently adapted a novel molecular biology technique to differentiate 16S rRNA gene sequences from different mycoplasma strains isolated from California sea lions (Zalophus californianus). This technique involves polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE) and automated DNA sequencing.