Development Of Method Of Detection HIV With Chemically Synthesized Polypeptide Antigen And Exploration Of Association Between CD4C868T Single Nucleotide Polymorphisms And HIV-â…  Susceptibility In Guangxi Population

Objectives1. Coating polystyrene plates with mixed polypeptide antigen including HIV Gp41, Gp120, Gp36and p24, and prepare diagnostic kit for HIV antibodies.2. Conducting performance Verification of this diagnostic kit for HIV antibodies.3. To explore the association between CD4C868T single nucleotide polymorphisms and HIV-Ⅰ susceptibility in Guangxi population.MethodsImproved methods of NalO4was used to labeled antigen with horseradishperoxidase (HRP-Ag) and polystyrene plates were coated with differentconcentrations of mixed polypeptide antigen, checkerboard titration was used toselect the appropriate concentration of mixed antigen and appropriateconcentration of HRP-Ag. The detection system was optimized withantigen-antibody reaction time, HRP-Ag reaction time and substrate reactiontime, and the sensitivity and specificity of the diagnostic kit were conducted. Polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) strategy and DNA sequencing methods were used to analyzepolymorphisms of CD4C868T (rs28919570C/T) in101patients with HIV-Ⅰ andage, sex-matched102healthy controls. χ2test was used to exam the differencesof genotype, allele, between the HIV-Ⅰ group and control group, logisticregression was performed to analysis odds ratio (Odds ratios, OR) with95%CI(Confidence Intervals, CI), adjusted for sex and age by the logistic regressionmodel, analysis the association between SNPs of polymorphisms and HIV-Ⅰsusceptibility in Guangxi population. SPSS16.0was used to conduct all thestatistics.Results1. The appropriate concentration of mixed antigen coated in polystyreneplates is5ug/ml, and the appropriate concentration of HRP-Ag is1:160.2. After optimizing the detection system, the antigen-antibody reaction timeis60min, the HRP-Ag reaction time is45min and the substrate reaction time is15min, the sensitivity and specificity of the diagnostic kit is97.2%and98.6%,respectively. The quality of the main active ingredient of the diagnostic kit arestable after be stored in25℃for a week or in4℃for one or three months.3. NO statistically significant was found for gender and age between HIV-Ⅰgroup and control group (P＞0.05). Genotype of rs28919570C/T in HIV-Ⅰgroup and control group were consistent with Hardy-weinberg equilibrium, andthe samples detected were representative and comparable.4. Three genotypes with CC, CT and TT were observed in rs28919570C/T,and no statistically significant was found for the frequency distribution betweenthe two groups （P＞0.05）. Comparing with CC genotype, either CT genotype or TT genotype was not associated with susceptibility to HIV-Ⅰ （P＞0.05）.5. The distribution of the rs28919570C/T SNPs in the control group weresimilar to the findings from the Han Chinese in Beijing, Japanese in Tokyo andGujarati Indians in Houston, but there were differences when compared with theAfrican ancestry in Southwest USA and Yoruban in Ibadan.6. Both in the male and female population, the three SNPs ofrs28919570C/T were not associated with susceptibility to HIV-Ⅰ（P＞0.05）.ConclusionsIt is feasible that diagnose HIV infection with mixed polypeptide antigencoated in polystyrene plates with high sensitivity and specificity. But noassociation between rs28919570C/T SNPs and HIV-Ⅰ susceptibility was foundin Guangxi population. Further research need to be conducted to confirm ourconclusions.