[Rapid centrifugation assay standarization for dengue virus isolation].

Dengue virus isolation remains a critical step in diagnostic and research workflows. Standard methods rely on tube-based cell culture techniques, which can be time-consuming and labor-intensive. Prior research has shown that centrifugation can enhance viral attachment to cells, but specific parameters for dengue virus remain underexplored. No prior work had resolved the optimal conditions for plate centrifugation in dengue virus isolation. This gap motivated the need for a standardized, rapid method. The study aimed to address this by evaluating centrifugation parameters. The focus was on improving isolation efficiency and reducing processing time. The goal was to provide a reproducible protocol for clinical and research settings. The study builds on existing centrifugation-based viral isolation strategies.

This study aimed to standardize a plate centrifugation method for dengue virus isolation. The specific problem was the inefficiency of conventional tube culture techniques. The motivation was to reduce the time and labor required for virus isolation. The study tested parameters such as centrifugation speed, inoculum volume, and sera dilution. The goal was to optimize these variables for maximum sensitivity and speed. The method was evaluated using clinical samples and reference viral strains. The researchers sought to validate the method against existing protocols. The study aimed to provide a practical and rapid alternative for dengue virus isolation.

C6/36-HT cells were used as the primary cell line for the experiments. The researchers tested various centrifugation speeds and durations. Inoculum volumes were adjusted to determine optimal conditions. Sera dilution ratios were evaluated for their impact on isolation. Incubation times were varied to assess virus replication efficiency. Indirect immunofluorescent tests were used to confirm viral isolation. Monoclonal antibodies were employed for serotype identification. The method was compared to the conventional tube culture technique. The study focused on reproducibility and sensitivity of the new method.

The optimal centrifugation speed was determined to be 1600 rpm for 30 minutes. An inoculum volume of 200 μL was found to be most effective. Sera dilution at 1/20 improved isolation sensitivity. The method achieved a sensitivity of 95.5% after five days post-inoculation. The plate centrifugation method outperformed the conventional tube culture. The time required for isolation was significantly reduced. The method was tested on 22 positive serum samples and four reference serotypes. The results showed consistent performance across all tested samples. The study confirmed the method's reliability and reproducibility.

The plate centrifugation method improved dengue virus isolation efficiency. The method reduced the time required for virus isolation. The optimized parameters included 1600 rpm, 200 μL inoculum, and 1/20 sera dilution. The sensitivity of 95.5% after five days was a key finding. The method was validated against conventional tube culture techniques. The study supports the use of plate centrifugation in clinical and research settings. The authors suggest that this method can enhance diagnostic workflows. The findings align with the goal of improving dengue virus isolation protocols.