| Objective: Through the study of HIV/AIDS cases in Yunnan Province from 2014 to2019,the distribution characteristics of HIV-1 strain subtypes and demographically related genotypes in Yunnan Province were described,the structural gene mutation,phylogenetic and amino acid variation of HIV-1 strains in Yunnan Province were analyzed,the endemic areas and time of Shuoyuan strains were explored;and new recombinant strains were sought.Method: Blood samples and epidemiological information were collected from 2800 HIV/AIDS patients in 16 prefectures(cities)of Yunnan Province from 2014 to 2019,the overall distribution of2014-2019 subtypes in Yunnan Province was described and the amino acid polymorphisms in the pol region were analyzed using the online analysis tool entropy;200 cases were randomly selected and their viral genomes were amplified and sequenced by qualitative RT-PCR.After sequence proofreading,the strain subtypes were determined using Genotyping,jp HMM,Mega6.06 and the online analysis tool BLAST,and the sequences were analyzed for amino acid variations to identify unique recombinant strains.Result:1.2800 samples from HIV/AIDS patients in Yunnan Province from 2014 to 2019 were amplified,of which 2570 samples were successfully typed,and the subtype accounting for the most proportion after typing was CRF08 BC subtype 54.5%,followed by CRF07 BC subtype 14.6%,and CRF01 AE subtype 14.2%.2.The distribution of subtypes in Yunnan Province showed obvious regionality,mainly in the eastern,central and western parts of CRF08BC;mainly in the northwest part of Yunnan Province,mainly in the western Baoshan and Nujiang areas of CRF01 AE,and mainly in the Dehong area of URFs.3.There were a total of 38 amino acid compositions of CRF08 BC subtypes with statistical differences in 2014-2019,of which 20 amino acid compositions showed a trend of increasing polymorphism over time;there were a total of 56 amino acid compositions of CRF01 AE subtypes with statistical differences in 2014-2019,of which21 amino acid compositions showed a trend of increasing polymorphism over time; there were a total of 51 amino acid compositions of CRF07 BC subtypes with statistical differences in 2014-2019,of which 21 amino acid compositions showed a trend of increasing polymorphism over time.4.Through the analysis of 132 nearly full-length sequences,CRF08 BC subtypes in our province formed four transmission clusters,of which the occurrence frequencies of characteristic amino acids in two transmission clusters were different to varying degrees,and the composition of characteristic amino acids in 35 sites changed.5.The horizontal gene distances of the three main CRFs in Yunnan Province were as follows: CRF01 AE > CRF07 BC > CRF08BC;in the CRF08 BC subtype,the average gene distance from large to small was env > pol > NFLG > gag;in the CRF01 AE subtype and CRF07 BC subtype,the average gene distance from large to small was env >NFLG > gag > pol.6.The selection pressure of the three main structural genes was negative selection pressure,and the selection pressure of CRF08 BC subtype and CRF01 AE subtype from largest to smallest was env > pol > gag;the selection pressure of CRF01 BC type was env > gag > pol.7.The unique recombinant strains in our province are mainly recombined in the form of subtypes B(B ’)and C,or in the form of subtypes BC and 01 AE for secondgeneration recombination.Conclusion: The subtype distribution of HIV/AIDS patients in Yunnan Province from2014 to 2019 has great changes in geographical distribution,transmission population and infection route compared with domestic and previous studies.Through the study on the near-full-length sequence,it is found that even the same subtype will form different epidemic trends in the same region.The study on the gene distance and selection pressure of different structural genes of different subtypes shows that CRF01 AE has richer amino acid polymorphism in Yunnan Province,with more advantages in epidemic.The molecular epidemiological changes of HIV-1 in our province should be closely monitored. |
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