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150 RFU 150 RFU Gabriele Savoldi: [email protected] Copan CPA200™ Semi-Automated Card Puncher Version 2 Performance Gabriele Savoldi 1 , Giorgio Triva 1 , Annalisa Gervasoni 2 and Santina Castriciano 2 , 1 Copan NewLab Engineering, Brescia Italy. 2 Copan Italia Spa, Brescia Italy Introduction The CPA200™ is a semi-automated punch system designed to process human blood or saliva samples, spotted on filter paper or cards, in forensic/genetics laboratories. CPA200™ offers punching accuracy and delivering of each punch into micro-wells plates, flexibility to process different card shapes and dimensions, samples traceability using barcode reader and camera control and integration into laboratory SOP and LIMS. After releasing CPA200™ in 2015, Copan NewLab Engineering developed the CPA200™ Version 2 (V2), currently available on the market. The CPA200™ V2 was developed after collecting all the requirements received from North America, Europe and Asia end-users. New features were added like extra punch of already processed cards, extra cleaning strike, resume protocol after interruption, customizable number of cleaning strikes, plate layout cloning, automatic export of reports, renewed graphic software interface, speed performance improvement and better compatibility with different brands of 96-wells plates. Objective The objective of this study was to demonstrate that the CPA200™ V2 can generate punches from saliva or blood samples deposited on cards or collector devices, to obtain human DNA profiling while preventing cross-contaminations and DNA carryover between wells/samples. Methods In this study, saliva and blood samples from different donors were spotted on Whatman® FTA® cards (FTAC), Copan NUCLEIC-CARD™ Color (NCC) for saliva, Copan NUCLEIC-CARD™ White (NCW) for blood and Bode Buccal DNA Collector (BBC) for saliva. Five 96- wells microplates were filled with punches performed by the CPA200™ following a ½ zebra + ½ checkerboard pattern, composed of negative control punches from blank cards alternated with punches from (1) NCC with saliva; (2) indicating FTAC with saliva; (3) NCW with blood, (4) non-indicating FTAC with blood, and transferred into empty wells; (5) BBC with saliva transferred into wells prefilled with Prep-n-Go™ Buffer. One cleaning punch was done from blank cards between consecutive samples to clean the puncher and avoid cross-contaminations. All punches were amplified with GlobalFiler® Express PCR Amplification Kit on Veriti® 96-well Thermal Cycler and fragments were analyzed on Applied Biosystems™ 3500 Genetic Analyzer. Data was analyzed with GeneMapper ID-X v1.4. Results When performing a zebra-checkerboard plate pattern with the CPA200™ V2 instrument, 100% of complete and reproducible DNA profiles were obtained from punches obtained from the three different sample card types spotted with saliva and blood samples. No cross- contamination or foreign alleles were detected in the samples. Additionally, no random carryover contamination was detected in any of the negative control punch. Overlay of 45 negative controls obtained when punching NCC with saliva. Overlay of 45 negative controls obtained when punching FTAC with saliva. Peak height obtained when punching NCC and FTAC with saliva.
