Serodia htlv 1 particle agglutination assay

Samples were initially screened at the Royal Darwin Hospital (RDH) or Institut Pasteur, Paris, using the Serodia HTLV-1 particle agglutination assay (Fujirebio, Japan) and Murex HTLV-I + II test kit (Murex Diagnostics, Dartford, UK). After November 2008, HTLV-1 screening at the RDH was with the Architect rHTLV-I/II assay. HTLV-1 serostatus was then confirmed by Western blot (HTLV Blot 2.4, MP Diagnostics) according to the kit manufacturer’s criteria at the National Serological Reference Laboratory (NRL), Melbourne, or Institut Pasteur, Paris. Attempts were made to confirm HTLV-1 infection for subjects with indeterminate Western blot results using HTLV-1 polymerase chain reaction (PCR) at the NRL. Primers and probes were designed to target a highly conserved 88 bp fragment of the gag gene in the p19 coding region of the Australo-Melanesian HTLV-1 subtype C. The sequence of the forward primer was AGT TCG GAG CTC AGG TCG AGA, the reverse primer was AGC AAG CAG GGT CAG GCA AAG and the probe was [6FAM]-GTCCGGCGCTCCCTTAGAGCC-[BHQ1] labeled with fluorophor FAM and Black Hole Quencher 1.

Initial screening tests were performed using the Serodia HTLV-1 particle agglutination assay (Fujirebio, Japan) or Architect rHTLV-I/II assay at the Royal Darwin Hospital, Northern Territory of Australia, (1458) or the Institut Pasteur, Paris (156). Positive samples were again tested using both the Serodia HTLV-1 particle agglutination assay and Murex HTLV-I+II test kit (Murex Diagnostics, Dartford, UK)(National Serological Reference Laboratory, Melbourne) or an indirect immunofluorescence assay (IFA) using an HTLV-1-transformed human T cell line (MT2)(Institut Pasteur). HTLV-1 serostatus was then confirmed by Western blot (HTLV Blot 2.4, MP Diagnostics) using stringent criteria for all samples for which screening tests were positive.