Dominant Drug-resistance Mutation Analysis On The HIV-1 Proviral DNA Of Infected Patients With Low Viral Load

Objective: To explore whether the proviral DNA carries dominant drug resistance mutations(DRMs)and DRMs distributions among HIV-1 infected individuals with low viral load(LVL)through HIV-1 proviral DNA drug resistance assays.To discuss the potential significance of HIV-1 proviral DNA drug resistance assays in clinical practice,therefore,to provide reference for clinicians to formulate and alter the antiretroviral therapy regimen for HIV-1 infected individuals with low viral load(LVL)and basis for predicting the prognosis of the disease.Materials and Methods: This research enrolled HIV-1 infected individuals who had received antiretroviral therapy more than 6 mouths in the HIV clinic of Changsha first hospital from May 2020 to May 2021.Their plasma RNA viral load(VL)was measured at 50 ～ 1000 copies /ml for two consecutive times in the baseline and the sixth mouth.We use nucleic acid extraction purification reagent(Hailite)to extract HIV-1 DNA,the fragment of HIV-1 PR/RT and IN genome was amplified by polymerase chain reaction(PCR)through In-house HIV-1 genotypic drug resistance assays.Stanford University Program HIV drug resistance database program(9.0version)and other tools were used to analyze HIV-1 sequence.Data processing and descriptive analysis were completed by Excel 2007.Results: 26 pol gene sequences were successfully obtained,we analyzed their HIV-1 proviral DNA dominant DRMs.1 case(3.84%)carried the dominant DRMs of nucleoside reverse transcriptase inhibitors(NRTIs),4 cases(15.38%)carried the dominant DRMs of non-nucleoside reverse transcriptase inhibitors(NNRTIs),1 case(3.84%)carried the dominant DRMs of protease inhibitors(PIs),and 1 case(3.84%)carried the dominant DRMs of integrase inhibitors(INSTIs).We found several dominant DRMs occurred in NNRTIs.3 cases showed drug resistance above low-level,1 case of which showed low-level drug resistance to current antiretroviral drug.And the rest showed potential low-level drug resistance.The incidence of CRF01＿AE was 53.84%(14/26),and dominant DRMs occurred in protease,reverse transcriptase and integrase gene sites.The incidence of genotype CRF07＿BC、CRF55＿01B、B+C、CRF67＿01B、CRF08＿BC、B were 19.23%(5/26),7.69%(2/26),7.69%(2/26),3.84%(1/26),3.84%(1/26)and 3.84%(1/26)respectively.In NNRTI genetic region,the incidence of dominant DRMs of V106 L was the highest,which was 7.69%(2/26),followed by M230 ML,V179E and V179 D.The DRMs in NRTI、PI、INSTI genetic region were K70 KT 、M46MI、H51Y respectively,and the incidence of each was 3.84%(1/26).Conclusion: 1.In this study,Sanger sequencing was used to detect the dominant DNA sequence with the highest percentage of abundance among the total sequences,to detect the dominant DRMS that cannot be detected among HIV-1infected individuals with low viral load(LVL)when using HIV-1 RNA drug resistance assays.2.In this study,the subtype CRF01＿AE had the highest incidence among HIV-1 infected individuals with low viral load(LVL).