Real-Time Reverse Transcription-Polymerase Chain Reaction Quantification of Tumor Necrosis Factor Alpha Messenger in Human Leukocytes
Alaaeddine, N.; Claudette de Montigny; M'Bark Sadouk
Clinical Laboratory 57(9-10): 799-802
2011
ISSN/ISBN: 1433-6510 PMID: 22029200 Document Number: 14534
Background: Tumor necrosis factor (TNF)-α plays a significant role in the pathogenesis of several inflammatory conditions. Several studies confirmed high TNF-α plasma protein levels in such conditions and highlighted the utility of TNF-α as a biomarker for disease progression or response to biological therapy. We aimed to provide a novel molecular technique for the measurement of TNF-α. Methods: Quantitative assay for the measurement of TNF-α messenger ribonucleic acid (mRNA) was done using real time reverse transcription polymerase chain reaction (RT-PCR). Results: The test was specific, sensitive, and efficient and possessed a wide scale of linearity. Intrassay and interassay CVs were equivalent to those observed in enzyme-linked immunosorbent assay methods for plasmatic TNF-α. Data from 11 healthy adults recruited in an attempt to establish reference values showed an average of 109.4 ng of U937 RNA (range: 67.0 - 173.5). Circulating TNF-α protein was not detected by ELISA in any of the 11 subjects. Conclusions: : The test may be useful for clinical studies and in research investigating TNF-α gene expression both in physiologic and pathologic situations, although these pilot findings need to be confirmed through larger studies.
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