| Objective:To investigate the relationship between EBV-DNA,T lymphocyte subsets(CD3~+,CD4~+,CD8~+cells,CD4~+/CD8~+ratio),serum levels of IL-2,IL-6,IL-8,IL-10,IFN-?and TNF-?)and atypical lymphocytes in laboratory diagnosis of Epstein-Barr virus-associated Infectious Mononucleos(EBV-IM)is laboratory sensitivity and specificity,provide a theoretical basis for the laboratory diagnosis of EBV-IM in children.Methods:A total of 34 children with positive EBV-CA IgM antibodies and clinically diagnosed as IM were selected as the observation group(group A).EBV-CA-IgM positive non-IM twenty-one children were positive control group(group B).20 children with non-IM with positive EBV-CA-IgG and negative EBV-CA-Ig M were negative control group(group C).Real-time fluorescent quantitative PCR was used to detect the EBV-DNA load in peripheral blood lymphocytes of three groups at the same time.The levels of CD3~+,CD4~+,CD8~+and CD4~+/CD8~+of T lymphocyte subsets in peripheral blood were measured by flow cytometry.The expression of IL-2,IL-6,IL-8,IL-10,IFN-?and TNF-?were detected by enzyme linked immunosorbent assay(ELISA),and the proportion of peripheral blood lymphocytes,blood and liver function were detected.Combined with the main clinical symptoms of children with EBV-related IM,with a comprehensive analysis of statistical methods.Res?Lt:1.The EBV-DNA load:The positive rates of EBV-DNA in the three groups were 82.4%,38.1%and 5%respectively.The positive rate of nucleic acid load in group A was significantly higher than that in group B and C(x~2=11.246,P<0.05).The sensitivity of EBV-DNA load to the diagnosis of children with EBV-related IM was 82.4%,and the specificity was 61.9%,positive predictive value was 77.8%,negative predictive value was 68.4%.2.T lymphocyte subsets:The ratio(%)of CD3~+,CD8~+,CD4~+and CD4~+/CD8~+T cells in group A were(85.02±6.22),(62.33±12.09)and(15.50±7.84),and its in group B were(70.56±7.43),(29.37±10.46),(34.45±9.78)and(1.37±0.71)respectively,and its in group C were(71.03±8.56),(30.13±8.93),(32.34±8.97)and(1.22±0.60),respectively.The number of CD3~+and CD8~+T cells in group A was higher than the upper limit of the reference level and significantly higher than those in group B and group C,while the ratio of CD4~+and CD4~+/CD8~+in group A T cells was significantly decreased.The difference was statistically significant(P<0.05).While its between group B and group C was not statistically significant(P>0.05).The diagnostic sensitivities of CD3~+,CD8~+,CD4~+and CD4~+/CD8~+were 61.8%,94.1%,91.2%and94.1%respectively in children with EBV-related IM.The positive predictive values were 100%,95.2%,85.7%and 90.5%.The positive predictive values were 100%,97%,91.2%and 94.1%,respectively.The negative predictive values were 61.8%,91%,85.7%and 90.5%respectively.3.Cytokines:The positive rates of IL-2,IL-6,IL-8,IL-10,IFN-?and TNF-?in group A were(7.30±5.46)ng/mL,(63.09±63.73)pg/mL,(149.46±122.05)pg/mL,(39.21±33.24)pg/mL,(836.20±675.01)pg/mL and(85.99±68.12)pg/mL.Its in group B were(3.79±3.25)ng/mL,(35.06±44.91)pg/mL,(89.99±99.57)pg/mL,(26.02±19.50)pg/mL,(495.90±509.10)pg/mL and(46.32±45.72)pg/mL.Its in group C were(4.55±5.51)pg/mL,(36.98±34.10)pg/mL,(92.32±69.59)pg/mL,(31.96±15.70)pg/mL,(548.49±318.83)pg/mL and(37.06±13.77)pg/mL.The levels of IL-2 and TNF-?in group A and group B were significantly higher than those in control group(P<0.05),while there was no statistical difference between group B and group C(P>0.05).The other cytokines(IL-6,IL-8,IL-10 and IFN-?)group A compared with the control group(groupB andgroupC),and the difference between group B and group C was not statistically significant(P>0.05).ROC curve analysis of IL-2 and TNF-?showed that the area under the curve respectivelywas 0.773 and 0.793,The levels of IL-2 and TNF-?at the maximum Youden index were 2.833 ng/mL and 34.347 pg/mL.The diagnostic sensitivities of IL-2 and TNF-?to children with EBV-associated IM respectivelywere 94.1%and 97.1%,and its specificity was 57.1%and 61.9%.ROC curve analysis of other cytokines(IL-6,IL-8,IL-10 and IFN-?)gave the area under the curve of 0.640,0.689,0.664 and 0.681.4.Comparison of auxiliary examination res?Lts:A,B and C three groups of children with abnormal blood samples were 27,12 and 5 cases;liver function in children were 26,3 and 1 cases;atypical lymphocyte proportion(>10%)increased in children were 20,0 and 0 cases.The positive rate of abnormal lymph node,blood and liver function in group A were significantly higher than those in group B and group C(P<0.001).The proportion of atypical lymphocytes in children with EBVassociated IM diagnosis sensitivity was 58.8%;specificity was 100%.5.Joint indicator diagnosis:The specificity of EBV-DNA and CD3~+in tandem(both were positive)was 100%.The sensitivity o f EBV-DNA in parallel(positive or positive)with IL-2 and TNF-?increased to 100%.The sensitivity of different lymph nodes in parallel with CD8~+,CD4~+,CD4~+/CD8~+and TNF-?increased to 97.1%.Conclusion:1.T-cell subsets(CD8~+,CD4~+/CD8~+)were highly sensitive and specific for the diagnosis of EBV-associated infectious mononucleosis in children,accounting for more than 94%and more than 90%respectively,followed by EBV-DNA.2.IL-2 and TNF-?have a better sensitivity in the diagnosis of EBV-associated infectious mononucleosis in children.3.The diagnosis of EBV-DNA in respectivelyparallel with IL-2 and TNF-?,and different lymph nodesin respectively parallel with CD8~+,CD4~+,CD4~+/CD8~+and TNF-?,can improve the sensitivity of EBV-related infectious mononucleosis degree. |
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