ABSTRACT
Objective To compare the consistency of the results of clinical specimens between Tecan Freedom Evolyzer-2200 automated enzyme immunoassay analyzer and manual ELISA method,so as to explore more scientific and reliable test method.Methods Ninety samples were randomly selected from clinical and physical examination,the IMARK2 microplate reader and the instrument operating manual sample measurement was used to test hepatitis A,hepatitis B 5,hepatitis C,hepatitis E 2,TP and HIV,and the results were given correlation and linear regression analysis.Results For the test items of HBsAg,HBsAb,HBeAg,TP,HIV,the correlation coefficient was r2＞ 0.9,the data showed a good correlation,and for HAV-IgM,HBeAb,HBcAb,HCV,HEV-IgM,HEV-IgG,the correlation coefficient was 0.9＞ r 2 ＞ 0.6.Compared with manual method,the total coincidence rate of automated enzyme immunoassay analyzer was 96.9％ in 11 indexes of 90 samples.Conclusions Automatic and manual ELISA method has a good correlation.Automatic ELISA also greatly reduce the labor burden of laboratory personnel,improve efficiency,should be strengthened in clinical practice.
ABSTRACT
Objective To analyze the association between the polymorphism of 5509-5511delCAA in exon4 of Tim-1 gene and the susceptibility to rheumatoid arthritis(RA)in a group of Han population from Yunnan province.Methods The PCR-DNA se-quencing analysis was used to detect the polymorphism of 5509-5511delCAA in exon4 of Tim-1 gene in 196 RA Han nationality pa-tients and 190 individuals of healthy physical examination(control).The ELISA method and indirect immunofluorescence assay was adopted to detect the three related aotuantibodies of RA,including the rheumatoid factors(RF)and the anti-cyclic citrullinated pep-tide(Anti-CCP)and the anti-keratin antibodies(AKA).Results There were statistically significant differences in genotype and al-lele frequency of 5509-5511delCAA between the RA group and the control group(P 0.05 ),but the positive rate of AKA between different genotypes of 5509-5511delCAA in the RA group had statistically significant differences (P <0.05).Conclusion The 5509-5511delCAA site of Tim-1 exon4 has the polymorphic variation,the 5509-5511delCAA genotype maybe related with the genetic susceptibility to RA in Han population of Yunnan province,the different genotypes does not affect the expression of RF and the anti-CCP,but affects the expression of AKA.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.</p><p><b>METHODS</b>The specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.</p><p><b>RESULTS</b>TCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.</p><p><b>CONCLUSION</b>The idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.</p>
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Anti-Idiotypic , Blood , Allergy and Immunology , Base Sequence , Complementarity Determining Regions , Genetics , Allergy and Immunology , Epitopes , Genetics , Allergy and Immunology , HL-60 Cells , Lymphoma , Allergy and Immunology , Mice, Inbred BALB C , Molecular Sequence Data , Receptors, Antigen, T-Cell, alpha-beta , Genetics , Allergy and Immunology , Sequence Analysis, DNA , Tumor Cells, Cultured , Vaccines, DNA , Genetics , Allergy and ImmunologyABSTRACT
This study was undertaken to investigate the anti-lymphocytic malignacy immunologic effects induced by TCR idiotypic DNA vaccine on BALB/c mice. CEM lymphoma cell line and BALB/c mice were used as models. The rearrangement gene fragment coding TCR Vbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into the eukaryocytic expression vector pcDNA3 to be used as DNA vaccine. The experimental animals were immunized by intramuscular injection with DNA vaccine. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The specific anti-idiotypic cellular immunity was detected by CTL activity assay using MTT method. The results showed that specific anti-idiotypic antibody in the immunized mice sera could be found since four weeks after immunization and came to the peak of titer on the sixth week. Using IL-6 as immunological adjuvant could significantly increase the antibody titers. It was concluded that the TCR idiotypic DNA vaccine could induce effectively the specific anti-lymphoma idiotypic antibody in BALB/c mice. Using IL-6 as immunological adjuvant could significantly increase the antibody titers induced by idiotipic DNA vaccine.