Evaluating the thermostability of commercial fast real-time PCR master mixes

Exp Mol Pathol. 2012 Oct;93(2):261-3. doi: 10.1016/j.yexmp.2012.05.002. Epub 2012 May 10.

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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / analysis*
  • Developing Countries
  • Drug Stability
  • Hot Temperature*
  • Humans
  • Point-of-Care Systems
  • Reagent Kits, Diagnostic*
  • Real-Time Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / standards
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction / standards

Substances

  • Reagent Kits, Diagnostic
  • DNA