| Objectives:In this study, we analyzed the changes of oral metabolites in HIV-infected patients by GC-TOF-MS platform, its aims is to ascertain the relationship between HIV stage of infection (different CD4 count levels) and oral metabolomics biosignature .The goal of this study will try to identify metabolic markers of HIV infection and provide important information for disease diagnosis and control.Methods:HIV-infected people who had not received highly active antiretroviral therapy(HAART) were enrolled in the Third People's Hospital of Kunming from September 2016 to December 2016. We collected the subjects' basic information and oral fluid samples and also recorded CD4 counts. The oral fluid samples were analyzed by non-target metabolomics using GC-TOF-MS and the data were pretreated, analyzed by PCA, OPLS-DA, metabolites qualitative analysis and differential metabolites screening, metabolic pathway and hierarchical clustering to analyze the changes in oral metabolites between HIV-infected persons at different levels of CD4.Results:In this study, 90 oral liquid samples from 90 HIV-infected patients were collected.They were divided into three groups according to peripheral blood CD4 count level:group A (CD4 count <200/mm3) cases, group B (CD4 count 200-500/ mm3) and group C (CD4 count> 500/ mm3) and each group of 30 cases.The samples received were performed by GC-TOF-MS and analyzed by bioinformatics and statistical analysis.The main results are as follows:(1) PCA analysis results: different samples in group A and group B, group A and group C are mostly arranged in a compact manner, clustered clusters and it showed a better experiment repeatability. However different samples I group B and C are mostly arranged relatively dispersed and failed to accurately show a better experiment repeatability; The difference between group A and group B, group A and group C was different from each other and a clear division was made which showing a good specificity. The samples of group B and group C were overlapped in different experimental groups, indicating that the similarity between partial samples was high and the difference was small.(2 ) OPLS-DA analysis results: samples in A and B , A and C are separated from each other which can be made a clear division. It can be said that the distinguishing between two groups of samples is very significant and the sample basically all in 95%confidence interval that imply this data is worthy of further study. And the distinction of the samples for B, C two groups is not very significant, but it can make a division and the distinction is also very obvious. The sample is basically all in the 95%confidence interval and it is indicated that this data is also worthy of further study.(3 ) The results of qualitative metabolites, screening differential metabolite,metabolic pathway analysis and hierarchical clustering analysis: There were 116 different metabolites and 7 kinds of different metabolic pathways in group A and group B. The most important metabolic pathways were Galactose metabolism,Pentose phosphate pathway, Pyrimidine metabolism, Linoleic acid metabolism,Fructose and mannose metabolism. There were 133 different metabolites and 9 kinds of different metabolic pathways in group A and group C. The most important metabolic pathways were Galactose metabolism, Pentose phosphate pathway, Purine metabolism, Pyrimidine metabolism, Linoleic acid metabolism, Fructose and mannose metabolism. There were 34 different metabolites and 4 kinds of different metabolic pathways in group B and group C. The most important metabolic pathways were Galactose metabolism, Fructose and mannose metabolism, Purine metabolism.Conclusion:The GC-TOF-MS was used to analyze the changes of oral metabolites in HIV-infected patients. The study found that the metabolic differences in oral are various form CD4 count levels. The results of the study can provide a new scientific basis for discovering new markers and controlling disease of HIV infection. |
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