Validation of PCR thermocycler performance is crucial to obtain reliable results. In this study, infrared (IR) thermography was evaluated as a novel validation tool. After stabilisation, no significant difference in the temperatures recorded using thermography and a reference block-based system was found. By employing IR thermography information about the length of the time until temperature stabilisation in the sample could be obtained. This study shows the potential of using IR thermography for validation of thermocyclers.Keywords: PCR, thermocycler, thermal cycler, infrared thermography, validation 3 Accurate thermocyclers is a prerequisite for all PCR based applications including diagnostic PCR for foodborne pathogens. Differences in temperature performance, between brands and types of cyclers has been demonstrated to influence PCR performance (Kim et al. 2008;Saunders et al. 2001;Yang et al. 2005). Proper validation of the thermocycler performance is one factor limiting the spread of diagnostic PCR to end-users (Malorny et al. 2003). Validation is important to ensure proper and reproducible results from PCR analysis and when transferring an assay between different thermocycler models (Schoder et al. 2005). It can also reveal problems caused by poor thermocycler performance (Kim et al. 2008;Schoder et al. 2005).Ideally, a validation procedure should be able to measure the temperature in-sample simultaneously in all wells. Two main approaches has been proposed in the ISO/TS 20836 standard (Anonymous when experiencing problems it can be hard to identify the source of the problem and therefore it is feasible to supplement the biochemical validation procedure with a physical one where the actual temperature within each well during temperature cycling is measured (Anonymous 2005;Schoder et al. 2003;Schoder et al. 2005;Yang et al. 2005). Probes for measuring temperatures in-tube have been developed (Schoder et al. 2003;Yang et al. 2005). Alternatively, the temperature distribution in the heating block (not in the sample) can be recorded using temperature sensitive probes. Several commercial systems are available for this purpose (e.g. TAS, Quanta Biotech, Somerset, UK and DRIFTCON, GENO-tronics, Landgraaf, The Netherlands). However, heat transfer and distribution through the sample itself, not the block alone, is crucial. Substantial variation in the effective length of a temperature step after the sample has reached the set temperature has been reported (Kim et al. 4 2008). For sensitive assays, this might affect the outcome of PCR and there is a need to calibrate the block temperature with the performance of PCR.Infrared (IR) thermography is a technology to detect and visualize IR energy radiating from an object for displaying images of temperature measurement or temperature distribution (for a review see, (Meola and Carlomagno 2004). The equipment used to perform IR thermography is often referred to as an IR camera or thermocamera. IR thermography has been employed as a tool for non-destructive evalua...
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