| Introduction and Objective:In China,HIV-1 has many subtypes,sub-subtypes and super-infection,suggested a variety of gene in HIV-1 strain.Exploring the spread of HIV can help us to understand the source of the infection of the virus and the propa-gation path,to reduce the probability of HIV transmission to uninfected,to explore th e implementation of the various prevention measures.We conducted three cross-sectiona Istudies:1?The aim of this study was to analyze the distribution ofclusters and drug resistance of CRFO1_AE among newly diagnosed,treatment-naive HIV-infected teenagers and you ng adults in 3 major HIV-affected geographic regions of Guangxi Province,including the cities of Hezhou,Liuzhou,and Nanning.2?The aim of this study was to investigate the origin and to reconstitute evolutionary pathway of subtype B HIV-1 among MSM in China.HIV-1 subtype B sequences of ne wly diagnosed,HIV infected MSM in 15 provinces or cities of China from 2007 to 2014 were included.3?Based on Illumina next generation sequencing(NGS)technology,we discuss the recombinants,super-infection and explore influences among MSM.Methods:A serial cross-sectional study:1K Samples were sequentially collected from newly diagnosed HIV infected 16 to 25-y ear olds in these 3 regions from 2009 to 2013.Phylogenetic analyses were used to de termine HIV-1 subtypes and CRF01_AE clusters.2?At last,a total of 354 subtype B sequences sampling from 2007-2014 were analyzed.Phylogenetic analyses were used to determine HIV-1 B clusters.Phylogenetic and Baye sian molecular clock analyses were used to define the clusters of B strains among MS M in different regions,and to determine the origins and divergence of major epidemic strains in MSM population.3?Based on the NGS Illumina platform,established an assay which can detect recomb inants and super-infection among MSM.This study was approved by institutional review board at the National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention.All data were analyzed using SAS 9.2 software packages.The proportion of HIV subt ypes over time was assessed using the chi-square trend test.P values<0.05 were cons idered statistically significant.Results:1?A total of 216 sequences were obtained from CRFO1_AE strains,which accounted for 83.1%of the 260 genotyped samples,of which 36 were from Hezhou,147 from Liuzhou,and 33 in Nanning.Most(83.3%,180/216)were from heterosexuals.Based on phylogenetic analyses by the maximum likelihood method,5 distinct clusters(cluster 1-5)were identified with 213(98.6%)sequences.In Hezhou,88.9%(32/36)of CRFO1_AE infections were caused by cluster 2,and 11.1%(4/36)were caused by cluster 1.In Liuzhou,83.0%(122/147)of theCRFO1_AE strains were found in cluster 1,11.6%(17/147)from cluster2,1.4%(2/147)from cluster 3,2.7%(4/147)from cluster 4,and 0.7%(1/147)from cluster 5.The distribution of CRFO1_AE clusters was moreeven in Nanning than it was in the other 2 regions,with 18.2%(6/33)from cluster 1,36.3%(12/33)from cluster 2,9.1%(3/33)from cluster 3,18.2%(6/33)from cluster 4,and 12.1%(4/33)from cluster 5.2.Twelve distinct B clusters were identified in 332(93.8%)sequences.Of note,57.3%(203/354)of the sequences were distinctly grouped into a large cluster(cluster 1)and widely distributed in Beijing(74.4%,151/203),Shanghai(14.0%,28/203),Hebei(2.9%,6/203)and other 10 provinces/cities in China(8.9%,18/203).Moreover,the sequences from Beijing were statistically occupied the root place of cluster 1.For further understanding the genetic characteristics of strains in cluster 1,three Near Full Length Genome sequences(NFLGs)were successfully amplified from the near root samples.The estimated time of the most recent common ancestors(tMRCAs)of this cluster was 1981(95%HPD,1977 to 1985),which predated the emergence of B'CN*and B'YN3?Finally,377(90.6%,377/416)were successfully amplified.Based on the NGS,81.7%(308/377)of the samples within the fragment has only one quasispecies.18.3%(69/377)samples ofpol genes and env gene do not match.The use of nitriteinhalants and alcohol use was alarmingly prevalent among MSM which were associated with high-risks of HIV-1 infection.The rate of inter-recombination and intra-recombination was 6.90%(26/377)and 2.12%(8/377),while the rate of inter-super-infection and intra-super-infection was 7.96%(30/377)and 1.32%(5/377)respectively.Conclusions:1?Clusters 1 and 2 of CRFO1_AE strains were prevalent in Liuzhou and Hezhou,respectively.However,multiple CRFO1_AE clusters existed in Nanning.This can be partially explained by the high mobility of laborers in Nanning,the capital city of Guangxi.2?The subtype B strains among Chinese MSM are genetically diverse.Cluster 1 was not young and it spreads quickly in recent years.The biggest cluster(cluster 1)is a long-standing and independent phylogenetic cluster prevailing in Chinese MSM.It might be related to the migration of MSM from Beijing to other cities in China.3?Based on the NGS Illumina platform,established an assay which can detect recombinants and super-infection among MSM.At last the super-infection among MSM in Beijing was 9.28%. |
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