Establishment Of Preliminary Diagnosis Method For Multiple Sclerosis Through Anti-Nucleic Acid Antibodies Detection

Multiple Sclerosis(MS) is a common autoimmune disease occurs in a wide range of central nervous system. The pathogenesis of the disease is not clear, and not find a cure for the disease yet. But some studies have shown that, the antinucleic acid antibodies were found in serum and cerebrospinal fluid of MS patients, and the anti-nucleic acid antibodies gathered in the lesions. Thus, we use this particular phenomenon, plasmid and natural extracts RNA as antigen respectively, established indirect Enzyme Linked Immunosorbent Assay(ELISA) to I diagnose MS through detecting anti-double stranded DNA antibodies and anti-RNA antibodies.First, we use Myelin Oligodendrocyte Glycoprotein Peptide(MOG35-55) immunized C57 BL / 6 female mice to establish animal model of multiple sclerosisExperimental Autoimmune Encephalomyelitis(EAE) model. After we found hindlimb paralysis and other symptoms characteristic of EAE in the mice, we killed the mice and removed of brain tissue and spinal cord in mice completely, and determined animal models established successfully by using immunofluorescence staining and demyelinating.Secondly, we set up indirect ELISA methods to diagnose MS through detecting anti-double stranded DNA antibodies and anti-RNA antibody. The indirect ELISA for detection of anti-double stranded DNA antibody: the lowest detection limit: 0.16 ng/m L, the linear range of detection: 0.5 ng/m L-100 ng/m L; the indirect ELISA for detection of anti-RNA antibodies optimized conditions: natural RNA coating concentration: 20 μg/m L, 100-fold dilution of serum. By indirect ELISA methods described above has been established to analyze the anti-double stranded DNA antibodies and anti-RNA antibodies in the serum and cerebrospinal fluid of EAE mouse model(n = 15) and the control group(n = 15), the results showed that the anti-double stranded DNA antibodies and anti-RNA antibodies in the cerebrospinal fluid and serum of mice in groups of pathogenic(n = 15) are much more than the control group(n = 15)(p < 0.01); also using the indirect ELISA methods described above has been established to analyze the multiple sclerosis(n = 9) and the other five neurological diseases clinical serum samples(each n = 9) obtained from the patient’s in first hospital of Jilin University and the healthy volunteers(n = 9), results showed that the content of anti-double stranded DNA antibodies in the serum of MS patients is more than 300 ng/m L, and the content of anti-double stranded DNA antibodies in the other five neurological diseases and normal human serum is less than 50 ng/m L(p < 0.01); anti-RNA antibody in the clinical patient serum detected valu is more than 2.1 times the negative control, while the anti-RNA antibodies in the other five neurological diseases and normal serum detected value is consistent with the negative control(p > 0.05). So we got two high specificity and sensitivity detection methods. This study provides a method and experimental foundation for simple, rapid, accurate clinical diagnosis of MS.