Use of lyophilized reagents on cellulose membrane in a LAMP reaction to detect Lambda phage virus

Lyophilization is a technique used to store prepared reagents while reducing the risk of error or contamination. In this paper, this technique was used to preserve reagents for Loop-mediated isothermal amplification (LAMP), a molecular amplification technique to detect low copy numbers of nucleic acids using isothermal conditions. While previous research has used lyophilized master mix in LAMP reactions, studies have not yet investigated the outcomes of target amplification with LAMP using both master mix and primer lyophilized on interfaced cellulose membranes. In this study, we performed LAMP reactions targeting Lambda phage bacterial virus. The assay was performed in a plastic pouch on a hot plate using lyophilized reagents on cellulose membranes. This was compared with standard methods in an Eppendorf tube, and with reactions on a membrane in a tube. Gel electrophoresis was used to compare the end product for the experimental and control methods. This amplification of the target DNA region demonstrates that the lyophilized reagents worked effectively the same as the solution reagents. Researchers are currently using this specific technology to develop a point-of-care diagnostic device to test for SARSCoV-2. The practicality of lyophilization of reagents enable ease of use in potential point-of-care applications of molecular methods.