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Published on: September 16, 2017
Diagnostic methods for the measurement of human TNF-alpha in clinical laboratory
Alan Valaperti1, Zhe Li1, Maya Vonow-Eisenring1
1Department of Clinical Immunology, University Hospital Zurich, Gloriastrasse 23, 8091, Zurich, Switzerland.
Insights
Measuring Tumour Necrosis Factor alpha (TNF-α) in blood is crucial for assessing inflammation. However, current immunoassay methods lack standardization, leading to unreliable results and challenges in clinical interpretation.
Area of Science:
- Immunology
- Clinical Chemistry
- Biomarker Analysis
Background:
- Tumour Necrosis Factor alpha (TNF-α) measurement in peripheral blood is vital for diagnosing and monitoring inflammatory diseases.
- Accurate and reliable quantification of TNF-α is challenged by the lack of standardized analytical tools with high specificity, accuracy, and sensitivity.
- Existing immunoassays, while robust, are often not interchangeable, complicating clinical laboratory practice.
Purpose of the Study:
- To compare the performance of various immunoassays for measuring TNF-α in human blood.
- To evaluate the suitability of different analytical methods for routine clinical laboratory use.
- To assess factors affecting TNF-α measurement accuracy, including sample type, stability, and potential interferences.
Main Methods:
- Comparison of ten different immunoassays: two ELISA, four flow cytometric bead array (CBA), and four Luminex assays.
- Evaluation of assay performance based on pre-established validation criteria (specificity, accuracy, linearity, precision, stability, sensitivity).
- Assessment of sample stability in serum and plasma, and potential interference from anti-TNF-α biologics.
Main Results:
- Significant lack of correlation was observed between different immunoassay techniques, with some correlation noted within the same technique.
- Only a few of the evaluated assays met the pre-defined validation criteria for clinical use.
- Reference values differed significantly between serum and plasma from the same donor; serum was stable for one week at 4°C, while plasma required freezing.
- Relevant interference was detected from anti-TNF-α biologics, potentially causing false-negative results.
Conclusions:
- The measurement of TNF-α in human blood is not yet harmonized across different clinical laboratories and immunoassay platforms.
- Few available immunoassays meet the stringent validation criteria required for reliable clinical decision-making.
- Clinicians must carefully consider potential interferences, especially from anti-TNF-α therapies, when interpreting TNF-α results to avoid misdiagnosis.
Abstract:
Measurement of Tumour Necrosis Factor alpha (TNF-α) in peripheral blood is a useful tool to assess inflammatory responses in a large range of diseases. One of the major challenges for cytokine analysis is the availability of a proper analytical tool with high specificity, accuracy, linearity, precision, stability, and analytical sensitivity. Although available immunoassays are usually robust and reproducible, it is also true that they are not interchangeable. Two ELISA, four flow cytometric bead array (CBA) and four Luminex immunoassays were compared. Correlation between different techniques was almost absent, while some immunoassays based on the same technique showed significant correlation. Among the ten different assays evaluated, just few of them complied with the pre-established acceptance validation criteria. Interestingly, sera and plasma collected from the same healthy donor had significant different reference values. Samples stability was maintained in serum up to one week at four degrees, while plasma was stable only when it was frozen. Since several anti-inflammatory treatments are based on biologics targeting TNF-α (anti-TNF-α antibodies), potential interference with the immunoassays was tested and resulted relevant. This study shows that although each immunoassay presents benefits and drawbacks, just few assays are suitable for the measurement of TNF-α in clinical laboratories, demonstrating that, so far, the measurement of TNF-α in human blood is still not yet harmonised. In addition, we found that false negative results caused by anti-TNF-α treatments should be carefully considered for results interpretation.
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