Whatman FTA Card As A Tool For DNA Preservation And Rapid Diagnosis Of Bacteria Diseases In Aquatic Analysis

Abstract

Flinders Technology Associates (FTA) cards simplify sample storage, transport, and extraction, therefore, the cost and time for diagnosis reduce. The purpose of this research was to evaluate the efficacy of the Whatman® FTA® card in inactivating bacterial cells and the storage of nucleic acids from bacterial cells that infected aquatic animals. This study was designed with 2 experiments: the FTA cards and filter papers were applied with different assumed bacterial concentrations of Vibrio parahaemolyticus (101 , 103 , 106 , and 108 CFU/mL) and infected Litopenaeus vannamei. After adsorption to FTA® Cards at room temperature for 3 hours, V.parahaemolyticus was completely inactivated within 24h, 48h, and 72h. Bacteria from the FTA cards isolated on TCBS revealed that there were no live bacterial cells from pure bacteria and infected shrimp whereas the bacterial count for the filter papers increased significantly. The bacterial DNA was sufficiently confirmed by the PCR technique, which was used to detect AHPND disease focused on the gene of V. parahaemolyticus at different time conditions at 4ºC. Moreover, the bacterial nucleic acid was intact after storage for 3 months and was PCR amplifiable. Overall, the results indicated that the FTA card was suitable for the secure transport without contamination and storage of aquatic bacteria under different conditions of a microbiological laboratory, as well as for the detection of their nucleic acid over various storage times.