Incidence of hepatitis C virus RNA in anti-HCV negative plasma pools in Croatia

The risks of transmitting viral infection by blood and products derived fro m plasma have long been known and still remain an area of concern. Blood ba nks and transfusion centres are faced with the imminent introduction of nuc leic acid amplification testing (NAT) of plasma pools as used by the plasma industry. In this paper, we show a part of our results of a validation stu dy of an in-house method for routine polymerase chain reaction (PCR) screen ing for hepatitis C virus (HCV) RNA in plasma pools and the results of test ing 2718 anti-HCV negative plasma pools for the presence of HCV RNA. The Eu ropean Committee for Proprietary Medical Products (CPMP) recommended that f rom I July 1999, only batches derived from plasma pools tested and found no n-reactive for HCV RNA, using validated test methods of suitable sensitivit y and specificity, should be batch released by authorities. The quality and efficiency of NAT detection of HCV RNA is among others influenced by the e fficacy of RNA isolation, the primer selection and the use of control sampl es. Using modern molecular biology techniques (sensitive and specific in-ho use amplification methods for detection of HCV RNA and automated sequencing ), we analysed samples of plasma pools from different Croatian transfusion centres. By detection of HCV RNA in an NIBSC working reagent (genotype 3) a nd a Pelispy HCV RNA run control (genotype 1) we determined a high reproduc ibility and sensitivity (below 100 International Units (IU)/ml) for our in- house method. By direct sequencing PCR cDNAs we proved the specificity of t he test system and the possibility of determining the HCV genotype when the method was used for PCR screening of HCV RNA in single donations. Of 2718 anti-HCV negative plasma pools we have found that 2.1% were HCV RNA positiv e. Results of our investigation confirm the necessity of testing HCV RNA in plasma pools to further increase the safety of human plasma-derived drugs. (C) 2001 Elsevier Science Ltd. All rights reserved.