| Objective:We followed up a group of 40 people who were first diagnosed with HIV infection and initiated the treatment of highly active antiretroviral therapy(HAART)for 6 months.Detected the expression of total HIV-1 DNA and the absolute count of CD4+,CD8+T lymphocytes in the peripheral blood of these patients before and after the treatment;explored the effect of HAART on the HIV “viral reservoir” which is represented by total HIV-1 DNA and discovered the role it played on the immune function.At the same time,we analyzed the correlation between the capacity of “viral reservoir” and the immune reconstitution;pursued the relationship between the expression of total HIV-1 DNA in peripheral blood and the efficacy of the treatment.Then built the ROC curve,and calculated the best cutoff value,accuracy and specificity of total HIV-1 DNA as a marker and index to evaluate the efficacy of HAART.Method: Utilized the BLAST-X to choose the conservative sequence(433-1435)of HIV-1 genome,which is located in the HIV-1 gag-pol gene.Using the beta actin as the internal gene and designing related primers,we amplified this HIV-1 DNA sequence in peripheral blood by real-time fluorescent Quantitative PCR(q PCR)and detected the relative expression of it.What's more,we use the technique of flow cytometry to count the CD4+T lymphocytes and CD8+T lymphocytes in peripheral blood.The differences of relative expression of total HIV-1 DNA before and after treatment in peripheral blood were compared by paired t test,as well the differences of the absolute count of immunocytes.Pearson correlation coefficient was use to analyze the correlation between the relative expression of total HIV-1 DNA and T lymphocyte count in each period.Finally,the ROC curve related to HAART treatment was established by using the ?total HIV-1 DNA as the indicator.In addition,the best cutoff value of the ?total HIV-1 DNA was calculated for judging the efficacy or inefficacy of treatment,as well as the sensitivity and specificity of this value of cutoff.Results: After 6 months,the study ended up with 32 eligible respondents,excluding those who failed to follow up or who did not meet the study criteria.In this case,the relative expression of total HIV-1 DNA(?x ± s)before and after treatment was 1.47±0.88,0.87 ±0.79 respectively,t=4.99,P< 0.05.The CD4+T lymphocyte count before and after treatment was 344.00 ± 157.64/ul,343.13 ± 121.17/ul respectively,t=0.056,P=0.956;CD8+T lymphocyte count was 755.44 ± 362.52/ul,785.53± 344.57/ul respectively,t =-0.580,P=0.566.There was a significant difference in the relative expression of total HIV-1 DNA before and after treatment.However,the difference in T lymphocyte count was very small,and had no statistical significance.The change of each index variation after treatment was stood by ?total HIV-1 DNA,?CD4+TC and ?CD8+TC respectively.Analyzed the correlation between each two of the them,the respective Pearson correlation coefficient was: r?CD4+TC&?CD8+TC = 0.653,P < 0.05,meaning there was a significant positive correlation between the ?CD4+TC and ?CD8+TC at 0.01 level(Two-tailed);r?total HIV-1 DNA&?CD4+TC=-0.741,P<0.05;r?total HIV-1 DNA&?CD8+TC =-0.527,P=0.002.It shown that the ?total HIV-1 DNA was significantly negatively correlated with ?CD4+TC and ?CD8+TC at the level of 0.01(Two-tailed).The area under the ROC curve(AUC)of ?total HIV-1 DNA corresponding to HAART treatment effect was 0.839,P=0.005,the optimal threshold(i.e.,cutoff)was ?total HIV-1 DNA =0.34,with a sensitivity of 70.8% and specificity of 87.5%.Conclusion: the treatment of HAART has a certain attenuation effect on the "viral reservoir" in the organism,which cannot be completely eliminated in a short time.The change of the "virus reservoir" before and after treatment has obvious linear correlation with the increase and decrease of CD4+ and CD8+T cell count.Total HIV-1 DNA is a better marker with a high sensitivity and specificity for evaluation of HAART efficacy,and it has a longer timeliness than conventional viral load detection. |
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