title: Evaluating the thermostability of commercial fast real-time PCR master mixes.
abstract: There is an increasing need for affordable, thermostable PCR reagents that can be used for diagnostic testing in resource limited settings. The development of point-of-care devices in such settings is highly dependent on the availability and efficacy of thermostable reagents. Here, we assess the thermostability of commercial, intercalating dye-based real-time PCR master mixes. We show that several of these master mixes have thermostability and robust performance at 20°C, 40°C, and 45°C for 6, 4 and 2 weeks, respectively. However, none of the master mixes that we evaluated was able to withstand more than 1 month at 45°C incubation. Our findings have implications for clinical diagnostics in the developing world where cold-chain delivery of diagnostic assays is difficult to guarantee.
authors.affiliation: Department of Pathology, Geisel School of Medicine at Dartmouth, Dartmouth-Hitchcock Medical Center, Lebanon, NH 03756, USA.
authors.forename: Ahmad N
authors.initials: AN
authors.lastname: Abou Tayoun
authors.forename: Brian P
authors.initials: BP
authors.lastname: Ward
authors.forename: George
authors.initials: G
authors.lastname: Maltezos
authors.forename: Axel
authors.initials: A
authors.lastname: Scherer
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Tsongalis
mesh_terms: D004247:DNA / Q000032:analysis*; D003906:Developing Countries; D004355:Drug Stability; D006358:Hot Temperature; D006801:Humans; D019095:Point-of-Care Systems; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards
chemical_list: D011933:Reagent Kits, Diagnostic; D004247:DNA