Real Time PCR User

You are generating real-time PCR (diagnostic) results and would like to support these results with appropriate evidence like biological controls, and assurance of full trace ability and quality of the used equipment. We therefore strongly recommend the use of DRIFTCON for real-time applications. Depending on the internal validation schedule, as well as the type of investigations, and the impact and importance of the generated result, you typically use DRIFTCON on a daily, weekly or monthly base, and trace the drift of temperature performance of your cycler in time. Your kit manufacturer may strongly recommend the use of DRIFTCON. With DRIFTCON you are comparing if the cycler used to run your application, is suitable for this application.

Every cycler has temperature un-uniformities. Be aware of the fact that you may have individual “hot” and “cold” spots. These spots are normally not detected with biological controls, or by the use of internal sample controls. In that respect it is important to realize that the results of fluorescence signal, yield and melting curves are in direct relation with these temperatures. If, e.g. the difference would just be 1°C, the impact on the generated yield and fluorescence signal., would already be enormous.

Your kit manufacturer may recommend the regular use of a DRIFTCON system to exclude variations in results, generated by a lack of performance of the used cycler. A kit manufacturer may even require seeing the thermal DRIFTCON profile of the used cycler prior to the acceptance of kit claims.

Your instrument may be performing out of “instruments specs” provided by CYCLERtest. Please request latest specifications by synchronising to our general database.

Users of commercially available kits may request kit manufacturers to provide prescribe threshold set(s) for temperature accuracy and block uniformity of the kit and/or cycler. Governmental auditors may request a kit manufacturer and other regularly bodies to provide the temperature threshold setting for the supplied kit as well as the actual validation of these data in time (DRIFTCON driven).

Threshold settings are used to compare the temperature performance of the cycler against (pre) defined thresholds. A kit manufacturer is capable to design extremely detailed threshold settings. These threshold settings are used to check if the tested instrument is within (application) thresholds or not. The kit manufacturer should provide a clear definition, whether if the threshold settings are based on application or on instrument.

Instruments thresholds settings (one specific instrument and block type) are provided with the purchase of a DRIFTCON system. Other individual specific threshold settings may be purchased as software “ad on” (option). The provided specific instruments thresholds are extracted from the CYCLERtest global database, which contains measurements from the same brand and type of machine. The provide threshold values are mean values plus or minus 2 x the standard deviation of the values (equals 95% of the population) You may adjust these settings by generating different defined instruments threshold setting.

Although the application thresholds are much more difficult and time consuming to design, they are superior to instrument thresholds. In other words: one is comparing if the cycler used to run an application is suitable for this application. If an end user is using a commercialised (diagnostic) kit (PCR, LCR, Real-Time PCR, RT-PCR etc) he or she may request the temperature profile thresholds from the kit manufacturer. If the kit manufacturer does not have these thresholds available consider using and participating our discussion forum at the specific DRIFTCON website: www.driftcon.com or (if available) join one of the application (your application) threshold setting group.