| Objective:The study we designed aimed at understanding the features of HIV-1 genotype?subtype distribution of newly reported HIV-1 positive people from Zhaoyang district Zhaotong city in 2016, providing theoretical basis for the region subsequent study of HIV-1 propagation rules and trends. To reveal the background prevalence of primary drug-resistance strains among newly reported HIV-1 positive people (drug-naive treatment), and grasp the relationship between gene subtype and drug resistance, in order to provide a reference for the establishment of preferred treatment and provide scientific basis for better antiviral therapy.Methods:Adopted cross-sectional epidemiological investigation for people who newly reported with HIV-1 positive and living in Zhaoyang district Zhaotong city Yunnan province, 2016. Plasma samples were separated from anticoagulation whole blood,and nucleic acids (RNA) of HIV-1 were abstracted by RNA abstraction kits. Target DNA fragments (env and pol gene sequence fragment) amplified by RT-PCR and nested-PCR, and the PCR products were sequenced. Gene sequences were disposed and spliced by SeqMan software. Phylogenetic analysis was conducted by MEGA software and recombinant analysis were conducted by SimPlot software. Pol gene sequence fragment were submitted to Stanford university drug-resistant HIV Drug Database for drug resistance analysis. All the epidemiological characteristics and molecular epidemiological data from HIV-1 positive people were analysed by SPSS 17.0 software, the correlation maps were conducted by Epilnfo software .Results:1. A total of 182 HIV positive cases were newly reported in Zhaoyang district in 2016, including 115 male (63.2%) and 67 female (36.8%); These people aged over 50 years old accounted for the largest (42.9%) among all the HIV-1 positive people;most of them were Han people(165, 90.6%), and 127 people were married, which accounted 69.8% in all cases. The majority of them just only received primary education, and some of them even without the chance ,witch accounted 74.7%. The main infection transmission was heterosexual contact, which accounted 92.8%, and most of the HIV positive people had the history of the unmarried heterosexual behaviour, including 89 (48.9%) people with non-commercial unmarried heterosexual behaviour, and 45 (24.7%) people with commercial unmarried heterosexual behaviour.The average degree of CD4+T cell count was 357.2±204.2cells/ mm3, and 46 people among all cases accounted 25.3%, whose CD4+T cell count was below 200 cells/mm3 were AIDS patients. In addition,the proportion of recent infected with HIV was high and the detection rate was 16.5% (30/182). Among all the HIV-1 positive people,most of them came from Fenghuang street? Taiping street, Sayu town and Jiupu town.2. 157 pieces of effective HIV-1 target gene sequences were obtained and the success rate of nucleic acid extraction and amplification was 86.3% in the study. We detected CRF01-AE subtype, CRF01-B subtype, CRF07-BC subtype, CRF08-BC subtype and other new recombinants among the 157 pieces. Generally, CRF08-BC subtype strains were the predominant circulating HIV-1 strains, which accounted 72.0% (113/157), the followed was CRF07-BC subtype accounted17.2% (27/157).The HIV-1 genotype composition was different in the different routes of infection.There were 75.3% patients were CRF08-BC subtype among the people who get infected with HIV though heterosexual transmission; the proportion of CRF07-BC subtype and CRF08-BC subtype were equal among drug uses, that was 40.0%; 60.0%patients were CRF07-BC subtype and 40.0% patients were CRP-01B subtype among these people who get infected with HIV though homosexual transmission. CRF08-BC subtype cases distributed in all towns except Buga huizu town in Zhaoyang district.CRF07-BC subtype cases were mainly distributed in Taiping street and Sujiayuan town, CRF01-B subtype cases were distribution in the Jiupu town and Leju town.3. In the research, a total of 135 pieceses of HIV-pol sequecces were successfully amplified and sequenced. Through genotype drug resistance testing, we detected that 39 patients had drug resistance mutation and the mutation rate was 28.9%.Then 32 patients had different degree of drug resistance related to protease inhibitors(PIs),nucleoside reverse trancriptase inhibitors (NRTIs) and non-nucleoside reverse trancriptase inhibitors (NNRTIs), the prevalence of drug resistance was 23.7%. The primary drug resistance situation belongs to the highly popular (> 15%) in Zhaoyang district. And among recent infection, the prevalence of drug resistance was 35.0%(7/20). In addition, NNRTs resistant mutation is as high as 93.9% among the three different types of resistance mutations. We detected 1 patient due the area of PR gene I50V mutation caused low drug resistant related to DRV and moderate drug resistant related to LPV/r, and 1 patient due the area of RT gene L100V mutation caused potential low drug resistant related to ETR, low drug resistant related to RPV and moderate drug resistant related to EFV, NVP.4. We didn't find drug resistance among CRF01-AE, while in CRF01-B subtype strain and CRF08-BC subtype stain, the prevelence of drug resistance was 100.0%,32.3% respectively. The detection rate of HIV-1 drug-resistant is different in different genotypes and that was statistically significant (Fisher's ?2= 10.5, P = 0.017). In addition, in the study we found in PR area all drug resistanance mutations occurred in CRF08-BC subtype, the locus mutation rate of 32nd up to 27.3% (3/11). In RT area,90.2% (37/41) of the drug resistance mutations occur in CRF08-BC subtype, the locus mutation rate of 138nd up to 64.9% (24/37). The mutation frequency in CRF08-BC subtype was higher than other subtypes.Conclusions:1. HIV epidemic situation was serious in Zhaoyang district. These people aged over 50 years old account for high propotion among newly reported in 2016, and heterosexual communication as the main route of transmission, most of people were AIDS patients when dected with HIV-1 infected, and the detection ratio of rencent infection account for high propotion. The majority of HIV-1 positive cases came from the main and the urban-rural fringe such as Fenghuang street, Taiping street and Sayu town.2. HIV-1 genotype present a certain degree of diversity in Zhaoyang district,CRF08-BC subtype strain had become the dominant HIV-1 strains among newly reported HIV-1 positive cases, and the followed by CRF07 - BC sutype strain, the distribution of HIV-1 subtype is differentin in different transmission route. In this reseach, the real recombinanting characteristics of 5 pieces of HIV-1 recombinants stains subtype also depend on the analysis of overall length of HIV gene sequencing.3. The drug resistance mutation rate and the prevalence of drug resistance was high among newly reported treatment-naive patients in Zhaoyang district. In addition, the prevalence of HIV primary drug resistance maybe at a higher level in the province even the state. The drug resistance mutation in PR and PR of pol gene result in different degrees of drug resistance related to PIs, NRTIs and NNRTIs.4. The prevalence of drug resistance and the propotion of mutations is different in different gene subtypes. We didn't find drug resistance among CRF01-AE subtype strain, while in CRF01-B subtype stain and CRF08-BC subtype stain the incidence of drug resistance was high, and the frequency of mutation is higher in CRF08-BC subtype stain than other subtypes. |
PDF Full Text Request