Use of the ADVIA 120 for differentiating extracellular and intracellular hemoglobin

BACKGROUND: The use of cell-free hemoglobin (Hgb) solutions, such as Oxyglobin (Biopure Corp, Cambridge, MA, USA), as a blood substitute for the treatment of acute anemias is increasing in veterinary medicine. These solutions interfere with colorimetric tests, which do not discriminate between cellular Hgb (Hgb-cell) from the patient and extracellular Hgb (Hgb-delta) from the Oxyglobin, and therefore make the monitoring of anemia, based on Hgb concentration, difficult. The ADVIA 120 hematology analyzer (Bayer Diagnostics, Tarrytown, NY, USA) evaluates Hgb by 2 methods, a standard cyanmethemoglobin colorimetric method and flow cytometry, and therefore might provide the means to differentiate extracellular and intracellular Hgb. OBJECTIVE: The objective of this study was to determine the accuracy and precision of the ADVIA 120 in differentiating extracellular from intracellular Hgb. METHODS: Anticoagulated whole blood samples from 10 healthy dogs were analyzed in triplicate on the ADVIA 120. Hgb-delta concentration was determined by adding Oxyglobin (13 g/dL) to the whole blood samples at dilutions of 1:1, 1:2, 1:4, 1:8, 1:16, and 1:32. Hgb-cell and Hgb-total values were calculated and compared with actual values by linear regression. Analyses were done in triplicate and repeated 9 consecutive times to evaluate intra-assay precision of Hgb-total and Hgb-cell determinations. RESULTS: Correlation between Hgb values obtained by colorimetric (Hb-total) and flow cytometric (Hgb-cell) methods on whole blood samples was high (R(2) = .99; n = 10) with a slope of 0.96 and intercept of 0. Correlation between actual and predicted Hgb-cell values also was high (R(2) = .99), with a small positive bias (0.289 +/- 0.185; n = 60). Intra-assay precisions were high, with most coefficients of variation <2%. CONCLUSION: The ADVIA 120 is capable of differentiating Hgb-cell from Hgb-delta. The flow cytometric method is accurate and precise when compared with the cyanmethemoglobin method. A small bias between the results is unlikely to be clinically significant but may affect the ability of the ADVIA to differentiate small quantities (<0.3 g/dL) of Hgb-delta.     

Validation of a Commercially Available Immunoassay for the Measurement of Bovine Cardiac Troponin I

Background: Commercially available cardiac troponin I (cTnI) assays developed for use in humans have not yet been validated for use in cattle.
Hypotheses: The ADVIA Centaur TnI-Ultra immunoassay can be used for the detection of bovine cTnI. In healthy cattle, serum cTnI is undetectable or is present only in trace amounts.
Methods: Purified bovine cTnI and cTnI-free bovine serum were used for the evaluation of assay performance including intra- and inter-assay precision, sensitivity, interference, linearity, and recovery. Effects of storage at 23, 4, −20, and −80 °C for 2 days, and at −20 and −80 °C for 7 and 14 days and repeated freeze-thaw cycles on recovery of cTnI were analyzed. Serum cTnI concentrations in 30 healthy dairy cows were determined.
Results: Intra- and inter-assay precisions (mean ± SD) were 4.48 ± 2.26 and 13.36 ± 6.59%, respectively. The assay demonstrated linearity at 0.5, 2, 15, and 30 ng/mL cTnI. Mean recovery was 100.81, 85.26, 87.72, and 114.42%, respectively. Skeletal muscle homogenate added to serum of known cTnI concentration did not alter the concentration of the analyte ( P > .05). Concentration of cTnI significantly decreased when samples were stored at 4 and 23 °C for 2 days ( P < .05). Repeated freeze-thaw cycles and storage at −20 °C for 7 days had no significant influence on cTnI concentration ( P > .05). Serum cTnI concentration in healthy cattle was ≤0.03 ng/mL.
Conclusion and Clinical Importance: ADVIA Centaur can be used reliably for the detection of serum cTnI concentration in cattle.