Lyophilized Ready-to-Use Mix for the Real-Time Polymerase Chain Reaction Diagnosis

Real-time polymerase chain reaction (real-time PCR) brings a more efficient and accurate method for detecting and analyzing nucleic acids in hospitals and laboratories. To solve the inconvenience of PCR reagent delivery and storage via cold-chain transportation, a solid-state reagent with robust characteristics should be employed. In this report, a lyophilized mix containing all necessary components for real-time PCR and its production method was designed, and its stability was tested at different temperatures. Some cryoprotectants and carriers are required to protect the function of the enzyme and primer as much as possible and provide the cake structure. Formulations with polyhydroxy compounds were considered to have the potential for protecting the enzymatic microstructure and functionality of dried mixes during the whole manufacturing and cold-free storage. The final products with the most superior protective formulation containing trehalose, Ficoll 400, and gelatin were able to provide the totally same testing result and sensitivity as the freshly made mix after 300 days of storage at 45 °C and can be deduced to maintain the function at room temperature (22.5-25.5 °C) for about 2 years.