| BackgroundOsteosarcoma(OS)is a highly malignant tumor.Its onset is hidden and its progress is rapid.If not treated in time,OS will cause serious consequences such as amputation even death.The discovery of early detection biomarkers is of great significance for clinical diagnosis.Anti-tumor-associated-antigen(TAA)autoantibodies can stably exist in the blood of tumor patients,and its expression level can be improved and detected before clinical symptoms appear.And the detection method is convenient and easy to accept.Therefore,autoantibodies can be detected as a serological detection method for early diagnosis of OS.ObjectiveThe aim of this study was to evaluate diagnostic value of eight kinds of anti TAAs autoantibodies(ENO1,GAPDH,HSP27,HSP60,NPM1,PDLIM1,STMN1 and TPI1)in OS.After ELISA,the best cut-off value and the best panel of autoantibodies were selected.Finally,a panel of serological candidate TAAs suitable for early diagnosis of OS was established.Methods1.Sample collection: a total of 130 subjects were included in this study,including 51 new diagnosed OS cases,51 healthy controls and 28 osteochondroma patients as benign controls.2.Indirect enzyme-linked immunosorbent assay(ELISA)was used to detect the expression level of 8 kinds of anti-TAAs autoantibodies in all the subjects.At the same time,scatter plot and ROC curve were used to screen the differential expression of autoantibodies,and the diagnostic value was evaluated.3.The best cut-off value and the best antibody panel were selected by Youden’s index method,95 th percentile method and mean plus two standard deviation method.The diagnostic value of the best panel was evaluated.4.All the data were analyzed by SPSS software.The statistical analysis methods included Kruskal-Wallis H test and chi-square test.All the statistical tests were bilateral,and the test level α was 0.05.Results1.The expression levels of seven anti-TAAs autoantibodies(ENO1,GAPDH,HSP27,HSP60,PDLIM1,STMN1 and TPI1)in OS patients were significantly higher than those in normal control group(P < 0.05).The area under the curve(AUC)of these seven autoantibodies ranged from 0.617 to 0.716,and the AUC of anti-ENO1 autoantibody was 0.716,the largest.2.The value of single autoantibody in osteosarcoma diagnosis was low.When cut-off value was set by the Youden’s index method,the sensitivity of single autoantibody was 39.22%,and the sensitivity of diagnosis can be increased to 66.67%by the optimal panel.When the cut-off value was selected as the 95 th percentile of the normal control group,the sensitivity of single autoantibody was 27.45%,while the sensitivity of diagnosis could be increased to 60.78% by the optimal panel.When mean plus two standard deviation method was selected for cut-off value,the sensitivity of single autoantibody in diagnosing osteosarcoma was 25.49%,and the sensitivity of diagnosis could be increased to 52.94% by the optimal panel.3.The results showed that the optimal combination of TPI1,PDLIM1 and GAPDH is the best panel with the highest Youden’s index.The sensitivity,specificity,Youden’s index,agreement rate and Kappa value are 66.67%,78.73%,0.45,72.55%and 0.45,respectively.4.According to the results of chi-square test,the difference of antibody positive rate between OS group and normal control group was statistically significant.The difference of antibody positive rate between osteochondroma group and normal control group was statistically significant,but the difference of antibody positive rate between OS group and osteochondroma group was not statistically significant.Conclusions1.ENO1,GAPDH,HSP27,HSP60,PDLIM1,STMN1 and TPI1 can be used as potential markers in the diagnosis of OS.2.The diagnostic value of single autoantibody in OS is low;the combined detection of three anti-TAAs autoantibodies(TPI1,PDLIM1 and GAPDH)can improve the diagnostic value of early diagnosis of OS. |
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