| Objective Objective to evaluate the role and value of RF,anti-CCP antibody and anti-MCV antibody in the diagnosis of rheumatoid arthritis,and to evaluate the application value of serum free DNA in the diagnosis and treatment of rheumatoid arthritis.To further compare the correlation between these parameters and clinical parameters,and to explore their role in the treatment and prognosis of rheumatoid arthritis.Methods First Part Serum samples were collected from 46 RA patients and 42 healthy subjects,the detection content of each case RF,anti-CCP antibody and anti-MCV antibody were collected,complement C3,C4,ESR,neutrophil count,lymphocyte count,duration,number of arthritis and other clinical data,to assess the value of RF,anti-CCP antibody and anti-MCV antibody in the diagnosis and treatment of RA.Second Part Construction the standard curve of ALU gene standard plasmid,set up a method for the detection of SYBR Green real time fluorescence quantitative PCR.Design 115 bp,247bp specific primers of the amplification product length to the ALU reference gene.Quantitative detection of serum DNA concentration of 46 RA patients and 42 healthy subjects,integrity and calculated free DNA,combined with clinical and pathological data of patients,assessment of free DNA integrity in RA and its clinical value.Results First Part(1)The sensitivity and specificity of RF in the diagnosis of RA was 82.6%and 83.3%.The sensitivity and specificity of anti-CCP in the diagnosis of RA was 76.1%and 95.2%.The sensitivity and specificity of anti-MCV in the diagnosis of RA was 84.8%and 92.9%.In the single item detection,the sensitivity of anti-MCV antibody was the highest,and the specificity of anti-CCP antibody was the highest.(2)When using the anti-MCV/anti-CCP method combined detection,the highest Youden index was 0.816,the sensitivity was 93.5%,specificity was 88.1%.(3)When using(RF/anti-CCP)+anti-MCV combined detection method,the highest Youden index was 0.826,the sensitivity was 82.6%,specificity was 100%.(4)The positive rate of RF was correlated with CRP level and duration of disease(P<0.05),that is,the CRP level of RF positive group was higher than that of negative group,the positive group of RF positive group was shorter than that of negative group,suggesting that RF content may be related to RA activity.Second Part(1)The standard curve of ALU gene standard plasmid were successfully constructed,set up a method for the detection of SYBR Green real-time fluorescence quantitative PCR.(2)The difference of ALU115 gene and ALU247 gene between RA group and healthy control group was statistically significant(P<0.001).The RA group was significantly higher than the healthy control group,the difference between the two groups was statistically significant(P<0.01).(3)Free DNA positive rate associated with the levels of C3,NE and RF content(P<0.05),the free DNA levels of C3 positive group was lower than that of negative group;free DNA NE positive group was higher than the negative group;the free DNA positive group RF was lower than that in negative group.It is suggested that the content of free DNA may be related to the activity of RA.Conclusion1.RF,anti-CCP antibody and anti-MCV antibody have a certain diagnostic value for RA,and the sensitivity and specificity of RA diagnosis can be improved to a great extent when combined with two or three indexes.2.The content and integrity of serum free DNA have a high clinical value in the diagnosis of RA,and the sensitivity and specificity of ALU115 gene in the diagnosis of RA were higher than those of RF,such as RA.3.The content of RF in the DNA positive group was lower than that in the negative group,which suggested that the free DNA test might be used in the diagnosis of RF negative RA4.Detection of peripheral blood cell free DNA has broad prospects in clinical application for its advantages of convenient and simple operation,small and easy to get and carry out large-scale... |
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