Summary The ELT-800 WBC and three-part WBC screen have been evaluated. Technical assessment showed each to be satisfactory but in a small number of samples, the total WBC was found to be incorrect. Discrepancies were found in some cases of chronic lymphatic leukaemia, sickle cell disease and para-proteinaemia. The reasons for these differences are discussed. In the population studied, an accurate WBC screen was obtained on 87.3% of samples. Initial instrument WBC screen rejection occurred on 12.7% but in only 2.1% could no explanation be found to account for this.
Abstract Non-human primates, such as rhesus and cynomolgus monkey share a similar biochemistry and metabolism to humans. These animals are therefore crucial model systems in a number of areas of biomedical research. To simplify cytokine profiling we have developed a 28-plex magnetic luminex bead-based cytokine immunoassay for the analysis of rhesus and cynomolgus monkey samples. Validated sample types include serum, plasma, and conditioned tissue culture medium. Analytes include: EGF, eotaxin, FGF basic, G-CSF, GM-CSF, HGF, IFN-γ, IL-1β, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 p40/p70, IL-15, IL-17, I-TAC, MDC, MIF, MIG, MIP-1α, MIP-1β, MCP-1, RANTES, TNF-α, and VEGF. The utility of the assay is demonstrated by determining cytokine profiles obtained with LPS-stimulated or PMA plus A23187-stimulated peripheral blood mononuclear cells (PBMCs) from rhesus monkeys and cynomolgus monkeys. Linearity of dilution experiments with tissue culture medium and serum produce R2 values of 0.98 and greater, while sample recovery routinely falls between 80 and 120%. Intra-assay and inter-assay precision is ≤10% CV. This multiplex monkey cytokine assay provides a very powerful tool for addressing cytokine profiling in non-human primates. 28 cytokines can be simultaneously analyzed from a single sample greatly simplifying and cutting cost of monkey cytokine analysis.
The underestimation of WBC by the Ortho ELT800/WS cell analyser in some cases of chronic lymphatic leukaemia has been investigated. Incorrect WBC was obtained on 44% of samples. WBC, smear cell count, lymphocyte cell volume, stage of disease and chemotherapy were studied in both discrepant and non-discrepant cases. The evidence suggested that low lymphocyte cell volume was the major cause of the anomaly. The relationship of small lymphocyte cell volume to increased cell density is also discussed.
Abstract Multiplexing immunoassays have become a very powerful and economic way to measure multiple analytes from a single sample. This is especially important when sample volumes are limited, such as for rodent samples. Life Technologies has recently launched a set of luminex bead-based immunoassays that can conveniently quantify between 10 and 30 cytokines from human or mouse tissue culture, serum and plasma samples. These assays use magnetic beads which offer a convenient way to automate these assays using commercially available plate washers, such as BioTek ELx405 or Tecan HydroSpeed. The mouse analytes in these kits are: GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 p40/p70, TNF-α (10-plex), supplemented with FGF basic, IL-1α, IL-13, IL-17, IP-10, KC, MCP-1, MIG, MIP-1α and VEGF (mouse 20-plex). Human analytes are GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10 and TNF-α (10-plex), supplemented with IL-1RA, IL-2R, IL-7, IL-12 p40/p70, IL-13, IL-15, IL-17, IFN-α, MIP-1α, MIP-1β, IP-10, MIG, eotaxin, RANTES, MCP-1, VEGF, G-CSF, FGF basic and HGF (human 30-plex). All the kits have been extensively validated to include sample recoveries routinely between 80 and 120%, intra-assay and inter-assay precision of <10%, and linearity of dilution with tissue culture medium and serum with R2 values of 0.98 and greater. Benchmarking of the kits has been performed against our ELISA kits to ensure equal results between the two immunoassay platforms.
Abstract An anti‐A agglutinin was found in the serum of a patient of blood group A 1 rhesus positive. The unusual feature of this antibody was the need for the presence of borate ions before activity could be demonstrated. No association of the occurrence of the antibody with a clinical disease was found.
