The Alpha-defensins (AD) present in synovial fluid have been regarded as constituting the\nmost accurate periprosthetic joint infection (PJI) biomarker. The methods most commonly used for\nestimating AD as a biomarker are the qualitative Synovasure® PJI tests, based on the technique of\nlateral flow, and quantitative enzyme-linked immunosorbent assay (ELISA). Here, we propose a novel\ntest based on detecting Alpha-defensins in synovial fluid by high-performance liquid chromatography\n(HPLC). Synovial fluid was collected from 157 patients diagnosed with PJI, infectious arthritis (IA),\narthrosis, reactive arthritis, and rheumatoid arthritis. AD concentrations in the fluid were determined\nby HPLC, and these same samples were used for additional diagnostic analyses. The results were\nstatistically processed to calculate cutoff concentrations for PJI and IA. HPLC testing showed a\nsensitivity of 94% and a specificity of 92% for diagnosis of PJI, and a sensitivity of 97% and a specificity\nof 87% for diagnosis of IA. Using HPLC, we detected in synovial fluid a combination of three\nAlpha-defensins: human neutrophil peptides HNP1, HNP2, and HNP3. All measured AD concentration\nvalues shown in this work refer to the sum of these three individual concentrations. Our study shows\nthat the HPLC method meets the conditions for measuring precise concentrations of the sum of AD\nand can be recommended as part of a diagnostic array for PJI and IA diagnostics. By this method,\nwe have verified that higher levels of AD in synovial fluid can also be seen in rheumatoid illnesses,\ncrystal arthropathies, and reactive arthritis.
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