A novel rolling circle amplification assay to detect members of the family Anelloviridae in pigs and humans

The study describes a novel rolling circle amplification (RCA) assay to detect members of the family Anelloviridae from swine and human serum samples. The RCA was carried out using short anellovirus (AV)-specific primers based on a highly conserved region among available AV full-length genomes. Then, RCA products were used as templates to amplify full-length genomes with an AV-specific PCR. Amplification products were separated by agarose gel electrophoresis and full-length genomes were isolated based on the known size. With this novel AV-RCA/PCR approach it was possible to detect Torque teno sus virus 1 (TTSuV1) and Torque teno sus virus 2 (TTSuV2) in swine and many species of Torque teno virus (TTV) in human sera, which were previously tested negative by conventional PCRs. © 2011 Elsevier B.V.