Preparation of Multiplex Real-time PCR for diagnosis of Influenza virus and Human respiratory syncytial viruses (RSV) spreading in Thailand

Abstract

Background The preparation of the Multiplex Real-time RT PCR technique is crucial for the diagnosis of influenza virus and Respiratory Syncytial Virus (RSV), both of which play significant roles in respiratory disease outbreaks in Thailand. Objective This study aims to prepare the technique for diagnosing respiratory viral infections, allowing for the simultaneous detection of multiple targets. It enhances the diagnostic capabilities of AFRIMS to support outbreak investigations and prompt respiratory disease outbreaks in both military personnel and civilians in high-risk areas, ultimately reducing the loss of army personnel. Method The research process involved using RNA samples to detect the genetic material of influenza and RSV viruses, with results compared to those from the Gold Standard (Real-time RT PCR) method using 30 positive samples and 30 negative samples, totaling 60 samples. These were tested for respiratory pathogen genetic material using the FDT^TM Respiratory Pathogens 21 reagent kit, alongside quality controls with Positive Control and Negative Control, with results compared to those from standard Real-time RT PCR testing. Result The results showed that the Multiplex Real-time RT PCR technique developed provided results consistent with those from the standard Real-time RT PCR for all 60 samples. Conclusion / Discussion This indicates that the developed Multiplex Real-time RT PCR technique at the Virology Laboratory of AFRIMS can accurately and reliably diagnose respiratory viral infections. This technique reduces the time required for investigating respiratory viral infections and prepares for handling disease outbreak incidents, thus improving the capability to control outbreaks and supporting future public health plans in Thailand.