Evaluation of a method for monoclonal antibody HLA-B27 analysis with the CELL-DYN Sapphire haematology analyser

SUMMARY

Analysis of HLA-B27 is usually performed by flow cytometry using commercial single or two colour fluorescence reagents. The CELL-DYN Sapphire (CD-Sapphire) is a high-volume routine haematology analyser that allows cell population analysis by monoclonal antibody fluorochromes analogous to flow cytometry. In this study, in-house flow cytometry analysis (n = 96, HLA-B27, One Lambda) performed on routine patient samples was used as the comparison method for analysis of HLA-B27, One Lambda (n = 40) and HLA-B27/HLA-B7, Immunotech (n = 96) reagents on the CD-Sapphire. The One Lambda results agreed 100% with the comparison method and offered clear population discrimination. The Immunotech combination also had a high level of agreement, but interpretation was more complex because of the wider cross-reactivity of the ABC-m3 antibody with B7 and other HLA-B alleles. When analysing HLA-B27 with antibodies showing nonspecific reactivity, a cut-off staining level yielding high specificity should be chosen, as the primary diagnostic value of HLA-B27 is as a ‘rule-out’ test for ankylosing spondylitis. The CD-Sapphire incorporates automated sampling and lysis, and medical scientists familiar with the instrument would require little additional technical training to perform the analysis. The reduced preanalytical work and total turnaround time constitute an important step towards automation of HLA-B27 and similar simple high-volume flow cytometry analysis.