Abstract
Serum and plasma calprotectin concentration is shown to be elevated when neutrophils are activated, and may therefore be used as a marker for inflammatory diseases. A serum calprotectin immunoassay was developed based on calprotectin values observed in samples from the intensive care unit. The polyclonal avian antibodies were raised and affinity purified with calprotectin antigens. The performance was tested and it was observed that the assay was linear in the range 0.3–24.7 mg/L, the limit of quantitation was observed to be lower than 0.3 mg/L, no antigen excess was observed up to 54 mg/L, all CVs were lower than 1.8 % in the precision study, the calibration curve stability was longer than 6 weeks, and there was no significant interference detected for haemoglobin, intralipid or bilirubin. The serum calprotectin immunoassay presented in this paper performs well within the criteria carefully set from the limited clinical experience obtained in both serum and plasma. In addition it is commutable with Bühlmann MRP8/14 ELISA.
Highlights
In clinical practice, the diagnosis of a bacterial infection requiring antibiotic treatment is made from observation of clinical symptoms, supported by leukocyte counts and/or C-reactive protein (CRP) measurement
It has been shown that the neutrophil activation marker HNL/NGAL could distinguish between acute bacterial and viral infections with a sensitivity and specificity of more than 95 % [1]
A particle enhanced turbidimetric immunoassay are usually used as a random access test and the samples will be run continuously as they arrive at the laboratory which will contribute to reduced test turnaround times
Summary
The diagnosis of a bacterial infection requiring antibiotic treatment is made from observation of clinical symptoms, supported by leukocyte counts and/or C-reactive protein (CRP) measurement. It has been shown that the neutrophil activation marker HNL/NGAL could distinguish between acute bacterial and viral infections with a sensitivity and specificity of more than 95 % [1]. Sepsis patients may have low levels of neutrophils in the blood as the neutrophil activation causes the neutrophils to attach to the endothelium and leave the circulation This can be circumvented by using a protein marker for neutrophil activation rather than measuring the number of circulating neutrophils. A particle enhanced turbidimetric immunoassay are usually used as a random access test and the samples will be run continuously as they arrive at the laboratory which will contribute to reduced test turnaround times. Avian antibodies do not react with rheumatoid factors, human anti-mouse IgG antibodies (HAMA) or the human complement system These are all well-known causes of erroneous test results in tests based on mammalian antibodies. The aim of the present study was to develop and validate a particle enhanced turbidimetric immunoassay (PETIA) for the detection of human calprotectin in serum or plasma
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