Optimization Of One-Step Quantitative Reverse Transcription PCR Assay For Direct Detection Of Rnase P Gene Expression From Human Saliva

Abstract

In recent decades, the issues relating to improving and expanding some medical tests, especially diagnostic cost, implementation process, and supply chain management are becoming more and more interesting. To deal with these problems, we tend to apply an efficient method to replace common methods, which is the use of saliva as a diagnostic fluid. The main positive feature of our approach is to use saliva instead of nasopharyngeal or nasal swabs, which enables non-invasive sampling without the need for trained staff. This advancement makes this material a safer source because of the protection of healthcare workers from being inadvertently exposed to potentially infectious droplets. In addition, we shortened the testing process by (1) not using universal transfer medium at sample collection, (2) the isolation and purification process were skipped, and (3) testing specimens in just one-step quantitative reverse transcription PCR (RT-qPCR) assay. During the project, several downsides of running real-time PCR on saliva samples had been successfully overcome. Pretreating the specimen with temperature and additives could overcome some inhibitTheThese above arguments can prove that saliva can be identified as a potential biomarker, besides noninvasive alternatives to swabs large-scale testing. Therefore, RT-qPCR assay on saliva is a flexible and inexpensive option to help improve diagnostic capacity in clinical as well as cope with future pandemics.