Study On HIV-1 Genotype Resistance And Molecular Network In Kunming

Objective(s):The purpose of this study is to understand the distribution of HIV-1strains and transmissible drug resistance in Kunming,and explore the transmission mode of HIV-1 in Kunming that understanding the characteristics of the molecular network of drug-resistant strains,judging the potential transmission and intervening in high-risk infectious sources based on molecular network technology.It will lay a foundation for the early research of large-scale HIV-1 transmission network monitoring in the future,and provide a reference basis for the selection of antiviral treatment programs for HIV-infected people and the improvement of antiviral treatment effects.Methods:The current situation investigation was adopted.HIV infected persons newly diagnosed whom current address in Kunming in Yunnan Infectious Diseases Hospital from June 2020 to December 2021.Demographic data were collected,patient blood was collected and CD4~+T cell test and HIV virus load test were performed.The HIV-1 pol gene was amplified by reverse transcription polymerase chain reaction and sequenced,sorted and compared.The sample sequence was uploaded to the HIV drug resistance database of Stanford University for drug resistance analysis.The ML phylogenetic tree was constructed by using the GTR model of the Fasttree software.Identification of the molecular networks and information extraction were performed by using the Cluster Picker software.The TN93 model of Hy Phy software was used to calculate the gene distances for pairwise sequences within a molecular cluster.Cytoscape software will presents molecular network visualization.Results:1.A total of 528 study subjects meeting the inclusion criteria were collected from June 2020 to December 2021 in Kunming.The study found that men accounted for 80.1%,and the age was mainly concentrated in the young and middle-aged people aged 16-45.The ethnic group was mainly Han(81.3%),and the marital status was mainly unmarried(52.3%),and the overall level of education was relatively high,most of which were college or above(37.7%).The main form of transmission was sexual transmission,of which heterosexual transmission accounted for 49.8%and homosexual transmission accounted for 43.8%.The mean baseline CD4~+T lymphocytes was 336±200 cells/mm~3.2.The HIV-1 genotype was dominated by CRF 08＿BC which accounting for26.9%,followed by URFs(25.4%)and CRF01＿AE(18.6%),CRF07＿BC(16.7%),CRF55＿01B(5.9%),and 35 other subtypes in Kunming.The restructuring of URFs was B/C restructuring in 118 cases,12 cases of 01＿AE/B/C restructuring,01＿AE/C recombination in 4 cases.The distribution of HIV-1 gene subtypes in Kunming had statistical differences in gender,age,nationality,marital status,education level and transmission route(P<0.05).3.The study found that the transmissible drug resistance rate in Kunming was11.7%,the NNRTIs drug resistance rate was 7.4%,the NRTIs drug resistance rate was0.8%,the PIs drug resistance rate was 3.6%,the INSTIs drug resistance rate was 0.4%,the double drug resistance rate was 0.6%,and HIV infection with triple and quadruple drug resistance was not found.Analysis of factors affecting drug resistance found that the risk of CRF08＿BC vs CRF01＿AE drug resistance was 7.192x of CRF01＿AE.The most common drug-resistant mutation site was V179D/E,with a mutation rate of12.6%.4.In Kunming HIV-1 molecular network,160 sequences entered the network,with a network access rate of 30.3%.A total of 61 clusters were identified,including16 molecule clusters of CRF 01＿AE,13 molecule clusters of CRF08＿BC,12molecule clusters of URFs,9 molecule clusters of CRF07＿BC,6 molecule clusters of CRF55＿01B,2 molecule clusters of CRF118＿BC,1 molecule clusters of CRF64＿BCand 1 molecule cluster of CRF85＿BC.In the network,55.1%of the heterosexual transmitted male infected persons had a potential transmission relationship with other heterosexual transmitted male infected persons.In the MSM population(MSM),95.1%of MSM had potential transmission associations with each other,and 28.2%of MSM were linked to heterosexual male infected persons(29.0%).5.The results of the analysis of the influencing factors of HIV infected people who entering the molecular network in Kunming showed that URFs and CRF07＿BC was not easy to access the network than CRF 01＿AE.Baseline CD4~+T lymphocytes of 350～499/mm~3and≥500/mm~3were easier to enter the network than baseline CD4~+T lymphocytes of<200/mm~3.6.CRF08＿BC had four drug-resistant transmission clusters which were heterosexual transmission,including two drug-resistant transmission clusters of K103N,a drug-resistant transmission cluster of E138A+V179D and a drug-resistant transmission cluster of E138A.CRF55＿01B had a drug-resistant transmission cluster of E138G+V179E which through homosexual transmission.In addition,there were 8potential drug resistance transmission clusters that all at the V179D/E in the network,with sex transmission pathways.7.The analysis results of high-risk communicators molecular network of Kunming show that no influencing factors of HIV infected persons as high-risk communicators were found.It is necessary to explore the influencing factors of HIV infection as high-risk communicators in Kunming.Conclusion(s):1.In Kunming,the subtypes of HIV-1 circulating strains from 2020 to 2021changed greatly compared with previous studies,which significantly increased proportion of URFs and diversification of subtypes.2.The transmissible drug resistance rate had reached a moderate level.The changes of transmissible drug resistance should be closely monitored.3.In the molecular network,CRF01＿AE has the most molecular clusters,CRF55＿01B has the highest access rate,and the largest molecular cluster of URFs.It is noteworthy that the transmission routes of CRF01＿AE,CRF55＿01B and URFs were mainly homosexual transmission.4.Molecular network of resistant strains found five clusters of resistant transmission and 8 potential drug resistance transmission clusters.It was suggested that there was a risk of transmission of HIV-1 drug-resistant strains in Kunming,and the drug-resistant transmission cluster should be precisely intervened to prevent the prevalence of drug-resistant strains.