COMPARISON OF EXTRACTION METHODS FOR DETECTING THE HEMOLYSIN GENE IN PATHOGENIC Aeromonas hydrophila FROM TILAPIA
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Abstract
Aeromonas hydrophila, a bacterial pathogen found in tilapia, negatively affects aquatic animal production yields due to the challenges in controlling disease outbreaks caused by the lengthy diagnosis process. This study compared two DNA extraction methods for A. hydrophila, a causative agent of disease in tilapia: an extraction kit and the heat boiling method.
The objective was to simplify and efficiently detect A. hydrophila carrying the hemolysin gene (hlyA). PCR assay results showed that both DNA extraction methods successfully detected the 592 bp amplicon of the hlyA gene without additional PCR products. The minimum bacterial concentration for effective DNA extraction and hlyA gene detection was 10⁶ CFU/mL. The DNA concentrations obtained were 18.8 ng/µL and 17.7 ng/µL, with purity values of 1.84 and 1.66 for the extraction kit and heat boiling methods, respectively. These results demonstrated that both DNA extraction methods effectively detected A. hydrophila via the hlyA gene, indicating reliable identification. Furthermore, the PCR results can potentially be used for DNA sequencing and gene-specific primer verification, facilitating the future application of these simplified analytical techniques.
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