Evaluation of automated nucleic acid extraction devices for application inHCVNAT

BACKGROUND: To further improve the safety of the blood supply, various nati onal blood transfusion organizations presently use or are in the process of implementing routine HCV NAT in minipools. According to the Committee for Proprietary Medicinal Products (CPMP) of the European Union, the HCV NAT de tection limit of the assay should be 100 IU per mt (270 geq/mL) for testing initial plasma pools. Paul Ehrlich Institute (PEI) regulations stipulate t hat 5000 IU per mL (13,500 geq/mL) must be detected to calculate the amount contributed by individual donations composing the minipool. The sensitivit y for HCV RNA extraction achieved by three commercially available laborator y kits was compared. STUDY DESIGN AND METHODS: Nucleic acids from 1-in-3 serial dilutions of an HCV RNA run control (Pelispy, CLB) were extracted with three kits (Cobas Am plicor, Roche Diagnostic Systems; BioRobot 9604, Qiagen; and NucliSens Extr actor, Organon Teknika). HCV PCR of all extracts was performed using a seco nd-generation Cobas Amplicor HCV test and the Cobas Amplicor analyzer. RESULTS: The manual Cobas Amplicor, the BioRobot 9604, and the NucliSens Ex tractor setups allow a 95-percent HCV RNA detection limit of 129, 82, and 1 2 geq per mt, respectively. The maximal pool size for the manual Cobas Ampl icor, the BioRobot 9604, and the NucliSens Extractor kits that would still meet the PEI criteria for HCV NAT in minipools was calculated at 104, 164, and 1125 donations, respectively. CONCLUSION: All three HCV NAT kits evaluated meet the criteria set by CPMP and PEI. The highest sensitivity for HCV NAT screening can be achieved with the high-volume NucliSens Extractor method in combination with the Cobas A mplicor HCV v2.0 test on the Cobas Amplicor analyzer.