Personal Temperature Drift Monitoring System For Thermal Cycling Instruments Product Info

INTRODUCTION This brochure provides general and technical information about the DRIFTCON system and is intended for (potential) end-users as well as for CYCLERtest Service distributors and MTAS agents/partners. The DRIFTCON system is personally used for the daily, weekly or monthly monitoring of the thermal performance of thermal cyclers which are used for any kind of thermal cycling reactions e.g. the PCR amplification process THE PCR -PROCESS AND TEMPERATURE PARAMETERS Reaction and process parameters in thermal cycling amplifications are not the only criteria that affect the quality of the results. Most PCR protocols use three temperatures in the process, (91 C-95 C) denaturation step, (40 C-60 C) annealing step and (70 C-74 C) extension step. This process is performed on a so-called thermal cycler that achieves the temperature cycling. The instruments for this application need to have excellent qualities in: Reliability - performing temperature cycling without interruption and failures Robustness and durability Temperature Accuracy the average block temperature must be within a specified range of the set temperature Temperature Uniformity the well to well difference (variation) must not exceed certain limits Reproducibility consistence in achieving the target temperatures at the same time in each cycle It is essential that the sample block and the PCR reaction mixture reaches the protocol temperature reproducibly and accurately. The so-called temperature overshoots and undershoots will have effects on the reactions as well as on the results. Secondly, sample block temperature uniformity is an important parameter in the PCR process. If the sample block heats uniformly, the control samples running in parallel with the samples will report if the block performs adequately. Non-uniform heating as well as temperature non-uniformities at the hold temperatures can lead to negative results of the control samples while the real samples have positive results or vice versa, all leading to incorrect conclusions. All of these above-mentioned issues are important while performing PCR reactions (regular, quantitative, diagnostic, ) with multiple samples. TEMPERATURE VERIFICATION For verification of the temperature quality of PCR instruments sample blocks, several methods are available. Even though, a simple mercury thermometer can be used for these specific temperature measurements, but this type of sensor is not suitable for accurate temperature measurements due to its robustness and inaccuracy. Other commercially available hand-held temperature measurement instruments may have a higher accuracy, but the used sensors do not match this special application. Furthermore, it is only possible to measure static temperatures on certain block locations sequentially, using hand-held single-sensor instruments, while the PCR process is a dynamic process. Due to a lack of accurate, independent, dynamic, and multi-channel temperature monitoring and validation systems for thermal cyclers, GENO-tronics and CYCLERtest developed a temperature measurement system for end-users optimized for this application, namely DRIFTCON. The system consists of special designed hardware and temperature probes for performing extremely accurate, sensitive and dynamic multi-channel temperature measurements. The temperature sensors used with the DRIFTCON are precisely designed and manufactured with the same cone angle and shaped as the sample wells. This means that for every sample block type there is an individually designed temperature probe. High thermal conductive cement with special parameters is used for molding the sensor in the precise shape. The bare sensors are not commercially available and are specially customized to suit the highly sophisticated DRIFTCON applications. The sensors have a high accuracy and sensitivity, as well as an extremely low drift and variation over time. Important parameters and specifications of the measurement system are the accuracy and the repeatability of the system. It is generally acknowledged that, if one wants to verify the quality and performance of a certain instrument, the accuracy and the resolution of the measurement system used has to be at least three times better than the instrument to be verified. The repeatability, reproducibility and drift of the complete measurement system are also considered important parameters. The DRIFTCON is Copyright 2002 GENO-tronics BV. All rights reserved. Page 2 of 8

designed to match all of these challenging parameters (see the paragraph system specifications for the technical details). Figure I: example of a DRIFTCON temperature probe fixture DRIFTCON APPLICATIONS As mentioned in the previous paragraph, GENO-tronics and CYCLERtest (both located in The Netherlands), have developed a highly sophisticated temperature measurement and validation system for capturing and analyzing the temperature performance of thermal cycling equipment, especially instruments used for PCR reactions, cycle sequencing instruments, incubators etc. The DRIFTCON has unique features and capabilities and is suitable for personal temperature validation monitoring as well as temperature drift monitoring of all kind and brands of instruments in the market e.g. thermal cyclers manufactured by MJ Research Inc, Applied Biosystems (PE), Eppendorf, Biometra, MWG, Thermo Hybaid, etc. The system has been calibrated against the ITS-90 temperature scale (NIST standard) and verified by an independent institute, which is accredited by the Dutch Council of Accreditation. Another striking feature of the DRIFTCON is its mobility. It s a mobile, out of the box and ready to use temperature-monitoring system. Users of thermal cyclers can now benefit from the convenience of frequent validation and accurate temperature drift monitoring on site, without having the necessity to send out the thermal cycler to a utilized test-site for validation. DRIFTCON test results are analyzed and reported through an automated process, thereby minimizing user input operations and prohibiting any alteration of the acquired sample data. STANDARDIZATION Every temperature measurement, regardless of which type of thermal cycling instrument is tested, is performed using a standardized and prescribed temperature protocol. This enables the comparison of the results of every measurement performed and shows if the results of tested instrument deviate from the protocol standards. This standard protocol is designed to support all relevant temperatures related to instrument specifications and the PCR processes. Data and results from the measurements are collected in a central database and the instrument specific field performance is calculated. The field performance can differ from the cycler manufacturer specifications. DRIFTCON users can also participate in a global database network generated by CYCLERtest. Particularly, users can determine how their instrument behaves in accuracy, uniformity as well as drift-in time in comparison to the other global users of the same cycler type and brand which also participate in the database and are also users of DRIFTCON. Besides the use of the personal DRIFTCON System (16 temperature channels), it is strongly recommended to use the external CYCLERtest Temperature Validation Service (MTAS-16, MTAS- 32, MTAS-48) once a year or after every 250 runs. The measurements and results of this unique service use even more channels (up to 48 simultaneously) and assure temperature validation of the highest standards and accuracy currently available. This validation comes with a detailed 16 pages of report which includes full dynamic accuracy, spread, uniformities, heating/cooling rates, graphs, gradient testing, load testing, recommendations and comparisons between same instruments etc. Copyright 2002 GENO-tronics BV. All rights reserved. Page 3 of 8

THE DRIFTCON SYSTEM The DRIFTCON System consists of two parts, 1) a hardware module with the temperature probe fixture and 2) the DRIFTCON monitor and analysis software. The DRIFTCON software controls the DRIFTCON hardware module and captures and processes the temperature data from the hardware module. The temperature data of each individual channel is acquired, plotted in real-time to a graph and stored to an encrypted logfile. The software indicates the actual temperature of each channel (up to 16) and calculates the average temperature and the temperature spread. The DRIFTCON-Card provides measurement credits; for each measurement a credit is deducted from the card. The software also analyses the data real-time and calculates main instrument parameters such as accuracy, spread, ramp speeds and other relevant data. DRIFTCON SOFTWARE The DRIFTCON software application is designed to provide easy operation and functionality. A graphical menu gains access to the main functions and features of the DRIFTCON software. Some key-features of the software are: o Device Management o User Management o Run Trace-ability o Reports o Synchronize Most functions are supported by a wizard to guide users through the steps. The picture on the right shows an example of the Device Wizard which allows you to add new instruments to your personal database and delete instruments or edit instruments. Figure II: DRIFTCON Device Wizard If a user wants to add a new instrument into the database a selection of the type of instrument can be made out of a list of instruments available in the DRIFTCON software. Most common thermal cyclers with all relevant data, parameters and (field) specifications can be made available. This picture shows an example of the Device Selection menu of a DriftCon user with several instruments. Figure III: Manufacturer and device selection The depicted pictures are just two examples of DRIFTCON features. There are more sophisticated features and smart functions making user interaction friendlier and clearer. Copyright 2002 GENO-tronics BV. All rights reserved. Page 4 of 8

DRIFTCON RESULTS All measurements are stored in a run trace-ability database. Figure IV shows an example of the Run Trace-ability tool of the DRIFTCON software. To each measurement a unique test number is assigned. All relevant data such test date, cycler identification etc. is stored with the run trace-ability. Figure IV: Example of the Run Trace-ability of DRIFTCON software. Next figures show the Analysis Result of a specific measurement of a certain type of instrument. The results are calculated at several temperatures and at certain time positions. For each parameter (spread and accuracy) the measured value and specification value are calculated and displayed. The result of the measured value is also indicated. The figures below show results at a temperature of 90 C and after 30 seconds. Figure V: Hold Time and overshoot at 90 C Figure VI: Results at 90 C and after 30 seconds One of the strongest features of the DRIFTCON software is that for every temperature hold in the test protocol, parameters and data for each individual channel are calculated simultaneously. All results and collected data that are used in the reports cannot be modified, which is an important issue in GLP/GMP/FDA and other Quality Programs. Depending on the user operation level the thresholds are adjustable. DRIFTCON s default threshold settings are based on average field specifications (market results). A user can choose different temperature and time plots to be displayed after the measurements. Copyright 2002 GENO-tronics BV. All rights reserved. Page 5 of 8

The next two figures show the drift results of the thermal cycler selected above. The figure shows the overview of the accuracy and the uniformity of the instrument. It has been tested 26 times in total. After measurement 14 the performance of this cycler has substantially improved. It has been repaired at that particular time. Figure VII: Selection of instrument for DRIFT calculations Figure VIII: Typical DRIFTCON drift graph This picture shows a magnification of performance improvement started at DRIFTCON validation #14. Diagnostics users could use the instrument from that date on again! Figure IX: Magnification of typical DRIFTCON drift graph DRIFTCON REPORTS DRIFTCON results are presented in detailed and partly selectable reports and relevant information is displayed and generated automatically. DRIFTCON offers several types of reports including retrospective overview of accuracy, uniformity, instrument and complete run trace-ability of all the variables used during the test. Participation in the global instrument database enables the user to compare the accuracy and spread against field specifications, generated using data from other global Copyright 2002 GENO-tronics BV. All rights reserved. Page 6 of 8

users with the same instrument. The thresholds for different users are not displayed in the database but may be available in specific user groups, which is a subpart of the database set-up. Privacy is assured since only the field specifications can be downloaded from the database and no other traceable personal information. CYCLERtest and GENO-tronics policy is to have uniformity and recognizability in the layout of the reports so that it cannot be changed or modified DRIFTCON SYSTEM COMPONENTS The DRIFTCON system consists of the following components: DRIFTCON-16 hardware module Intelligent CyclerProbe Fixture (available in several models and formats) o 16 x 0.2ml: 16 temperature sensors for 0.2ml block format o 16 x 0.5ml: 16 temperature sensors for 0.5ml block format o 16 x 384 fit: 16 temperature sensors for 384 block format o Other special models available on request DRIFTCON SMART-card (10 acquisitions) Connection Cable DRIFTCON Software Robust, CyclerProbe Fixture box Optional: State-of the-art Notebook with DRIFTCON software pre-installed SYSTEM FEATURES Dynamic Temperature Monitoring (DTM) Static Temperature Verifications (STV) Multi-channel (up to 16) Intelligent temperature probes fixtures NIST-traceable, validated by Dutch Council for Accreditation Main supply 90-240VAC (50-60Hz) CE-mark SYSTEM SPECIFICATIONS Specifications are subject to change without prior notice Accuracy 1 : average channel value accuracy up to <0.1 C or better (NIST-traceable) Measurement resolution: 16bits Software sample rate/channel: 2Hz Measurement data range: 10 100 C (1. Temperature range 30.0-95.0 C) DRIFTCON APPLICATIONS GMP/GLP validations FDA regulations, validations Diagnostic quality programs Other quality program qualifications Protocol transfer and cycler transfer Instrument to instrument comparison Periodic performance validations Process problem solving Real-time PCR Instrument validation Biotechnology applications, Clinical laboratory QC PCR plate temperature uniformity For correct, reliable and accurate measurements, trained and certified personnel should only operate the system. Copyright 2002 GENO-tronics BV. All rights reserved. Page 7 of 9

THE CYCLERTEST SERVICE For more information about DRIFTCON System or the CYCLERtest Thermal Cycler Validation Service or MTAS-16, MTAS 32 and MTAS 48 systems, we recommend you to contact CYCLERtest BV, The Netherlands. See paragraph contact for more details. TERMINOLOGY Accuracy The average temperature of the sample block, measured in more then two sample well locations simultaneously, compared to a traceable standard of absolute temperature (NIST). Uniformity Sample wells in the sample block of the thermal cycler should heat uniformly relative to each other. Uniformity, spread or homogeneity is the highest measured temperature values minus the lowest temperature at the same time. Static Temperature Verification Measured temperatures of several locations in the sample block at a static temperature hold sequentially. Measurements are not taken at the same time. Dynamic Temperature Monitoring Monitoring the temperature continuously on 4 to 16 locations simultaneously during the temperature changing process. CONTACT Distribution and Service Research & Production Facilities CYCLERtest BV GENO-tronics BV PO Box 31122 Rötscherweg 61, Entrance C 6370 AC LANDGRAAF 6374 XW LANDGRAAF The Netherlands The Netherlands info@cyclertest.com www.cyclertest.com info@geno-tronics.com www.geno-tronics.com TRADEMARKS o DRIFTCON, DRIFTCON-Card, MTAS, MTAS-16, MTAS-32, MTAS-48, MTAS-96, and CyclerProbe are trademarks of GENO-tronics BV. o All other brand names are (registered) trademarks of the respective holders and manufacturers. o The Polymerase Chain Reaction Process (PCR ) is covered by patents owned by Hoffman-Laroche, Inc. (4.683.195, 4.683.202, 4.965.188) Copyright 2002 GENO-tronics BV. All rights reserved. Page 8 of 9