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Furthermore, MS analysis of individual serum samples is important for biomarker discovery because it helps validate the pattern of differential protein expression among all RA patients. However, this discrepancy can be eliminated through individual sample analysis. Nevertheless, when some patients have a high abundance of certain proteins, these protein expression patterns seem to represent all patients. Finally, it is difficult to control the data processes. Third, clinical data interpretation is challenging owing to the complexity of the status of RA patients. Second, individual MS analysis is costly and time-consuming. First, it is difficult to obtain an adequate volume of individual serum samples. However, it is important to analyze individual serum samples to reflect individual alterations in serum protein levels, but it is difficult to analyze individual serum samples.
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In most previous proteomics studies, blood samples were pooled for MS analysis. New protein biomarkers potentially detected through serum protein profiling are expected to represent various physiological changes in RA, other than inflammation and immunity. Furthermore, current diagnostic biomarkers reflect the status of inflammation and immunity among RA patients.
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If these new biomarkers could diagnose seronegative (RF- and anti-CCP-negative) RA patients, they could contribute to the accurate diagnosis and treatment of RA. Therefore, novel diagnostic biomarkers complementing the existing biomarkers, i.e., RF and anti-CCP, are required. Anti-CCP has higher ACCP positivity than RF positivity among RA patients however, the sensitivity and specificity of the two markers do not significantly differ. To improve the efficiency of RA diagnosis, anti-CCP is used with RF. For example, the sensitivity and specificity of RF are 60–90% and 85%, respectively. However, existing biomarkers have limitations concerning RA diagnosis. Therefore, CRP, RF, and anti-CCP, representing the inflammatory and immune response of RA, are diagnostic blood biomarkers. When serum peptides are citrullinated or subjected to other posttranslational modifications by various environmental stimuli, the altered peptides are presented to immune cells, including T cells as antigens, and antibodies such as anti-CCP are produced. Hence, autoantibodies are overproduced, leading to an increase in immunoglobulin M (RF) and anti-CCP in RA patients.
#Rheumatoid arthritis diagnosis criteria series#
Furthermore, a series of immune responses are triggered with an increase in inflammation. Activation of inflammation increases cytokine, chemokine, and inflammatory reactants such as C-reactive protein (CRP). The disease is initially characterized by an inflammatory response, followed by autoantibody activation and damage to the synovial membrane and joints. Other risk factors include environmental factors such as smoking, oral health, and diet. Initially, these genetic factors include gene polymorphisms, epigenetic factors including DNA methylation and histone acetylation, and complex factors. However, genetic factors account for 60% of the RA risk factors. Rheumatoid arthritis (RA) is an autoimmune disease with an unknown etiology. Therefore, among seronegative RA patients, a four-biomarker panel (AGT, SAA4, VDBP, and RBP4) can prevent false negatives and help diagnose RA accurately.
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Seven biomarker candidates were verified through multiple reaction monitoring-based quantitative analysis, of which angiotensinogen (AGT), serum amyloid A-4 protein (SAA4), vitamin D-binding protein (VDBP), and retinol-binding protein-4 (RBP4) had an area under the curve over 0.8, thus distinguishing RA patients, including seronegative (RF- and anti-CCP-negative) RA patients, from healthy controls. We performed statistical and functional analysis of differentially expressed proteins to identify biomarker candidates complementing conventional serological tests. We aimed to identify alternative biomarkers by serum protein profiling using LC-MS/MS. These serological factors are diagnostic markers of RA however, their sensitivity and specificity for prediction warrant improvement for an early and accurate diagnosis. Rheumatoid arthritis (RA) is an autoimmune disease of inflammatory joint damage, wherein C-reactive protein and autoantibodies including rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) are rapidly elevated.