Molecular epidemiology of HIV-1 in central region of Nepal Nirajan Bhusal 1,2, Navin Horthongkham 1, Niracha Athipanyasilp 1, Wannee Kantakamalakul 1, Ruengpung Sutthent 1 1 Department of Microbiology, Faculty of Medicine Siriraj hospital, Mahidol University, Thailand. 2 Department of Microbiology, Kathmandu University School of Medical Sciences, Kavre, Nepal

data sources:AIDSinfoonline.org Trend of global HIV Infection and Death

HIV Prevalence:

Subtypes and CRFs distribution in Asia:

Epidemic snapshot of HIV in Nepal

Subjects

Clinical characteristics of selected sample Sample I.DYear of Infection Location Risk GroupViral loadART status NP Nawalparasi HSR3,685NONE NP Chitwan HSR79,787AZT,3TC,NVP NP Nawalparasi HSR5,942NONE NP Chitwan IDU3,298AZT,3TC,NVP NP parsa MTCT31,940NONE NP Chitwan HSR10,413AZT,3TC,NVP NP Chitwan HSR5,942AZT,37C,NVP NP Chitwan HSR63,485NONE NP Chitwan HSR14,420AZT,3TC,NVP NP Gorkha HSR4,599AZT,3TC,EFV NP Chitwan HSR22,777AZT,3TC,NVP NP Makwanpur HSR< 40AZT,3TC,NVP NP Chitwan MTCT8,64,318d4T,3TC,EFV NP Chitwan IDU4,599AZT,3TC,NVP NP Chitwan IDU40,192NONE NP Chitwan HSR159,488AZT,3TC,NVP NP Chitwan IDU25,980AZT,3TC,NVP NP Chitwan MSM2,176NONE NP Gorkha HSR2,230NONE NP Chitwan MTCT6,067NONE NP Chitwan IDU51,218AZT,3TC,NVP NP Nawalparasi HSR86,103AZT,3TC,NVP NP Nawalparasi HSR7,314NONE NP Nawalparasi HSR9,649NONE NP Chitwan MTCT3,186NONE

Sample I.DYear of Infection Location Risk GroupViral loadART status NP Chitwan HSR9,649NONE NP Nawalparasi HSR21,253NONE NP Nawalparasi HSR32,235NONE NP Chitwan HSR44,285NONE NP Parsa HSR4,695 AZT,3TC,NVP NP Chitwan HSR60,480NONE NP Chitwan HSR3,535NONE NP Nawalparasi MSM21,265AZT,3TC,NVP NP Chitwan HSR5,545NONE NP Chitwan HSR<40AZT,3TC,EFV NP Nawalparasi MSM1,819NONE NP Nawalparasi MTCT9,625NONE NP Chitwan HSR5,807NONE NP Nawalparasi HSR31,794AZT,3TC,NVP NP Chitwan HSR2,612AZT,3TC,NVP NP Chitwan HSR159,488NONE NP Chitwan IDU25,980AZT,3TC,NVP NP Chitwan HSR2,176NONE NP Gorkha HSR2,230AZT,3TC,NVP NP Chitwan MSM6,067NONE NP Chitwan HSR51,218NONE NP Nawalparasi HSR86,103AZT,3TC,NVP NP Nawalparasi HSR7,314NONE NP Nawalparasi HSR9,649NONE NP Chitwan MTCT3,186NONE Clinical characteristics of selected sample

Siriraj hospital in bangkok

Ethical issues

Characteristics Total patientsART Naïve patientsART experienced n n n Number of patients1073 Male642 Female431 Transmission mode Heterosexual541 Homosexual110 MTCT211 IDU211 medianmin-maxmedian Age (years) CD4 + T cells Viral load (log copies/ml) –

Phylogenetic analysis of 10 HIV-1 nearly full length nucleotide sequences

9 samples : 11NP004,11NP014, 11NP016, 11NP065, 11NP076, 11NP084, 11NP086, 11NP091 and 11NP115 1 sample 11NP079

Fragment Start Position Uncertainty Region Start - End Breakpoint Interval Start - End Fragment End PositionFragment Subtype Position in the original sequence [pred_recombinati on], [recombination_incl_UR _and_BPI], [UR_and_BPI][pred_recombinati on][recombination_incl_UR _and_BPI][UR_and_BPI] N/A C D C D C N/A Position based on HXB2 numbering [pred_recombinat ion] [recombination_incl_UR _and_BPI] [UR_and_BPI]HXB2 numbering[pred_recombinat ion][recombination_incl_UR _and_BPI][UR_and_BPI]

Sample ID HIV diagnosis year Route of infection Plasma HIV-1 RNA level (copies/ml) HIV DRM PI NRTI NNRTIDrugs 11NP MTCT31,940--K103R Efavirenz 11NP HSR10,413--V179D Delavirdine 11NP HSR2,230--E138A Etravirine 11NP IDU51,218--K103R Efavirenz 11NP822010HSR32,325A71V-- Ritonavir 11NP HSR2,612A71T-- Indinavir 11NP MTCT159,297--V108I Efavirenz 11NP MSM58,782A71T-- Mutation pattern of amino acids and drug resistance

Subtype C NP015 one major mutation at position 82 one minor at 60 CRF-08BC NP008 three minor mutation at position 36,60 and 77 Position B-cons T V L E E M N L P G R W K P K M I G G I G G F I K V R Q Y D C-cons..... V S... K. R E NP D I D Q NP N D H Position B-cons A I G T V L V G P T P V N I I G R N L L T Q I G C T L N F C-cons M NP I NP I Amino acid polymorphism in different subtypes

Signature sequence pattern analysis NP003- HSRT C T R P S N N T R K S I I G P G Q T F Y NP131- MSMN.... N NP003- HSRA T G D I I G D I R Q A H C N I NP131- MSM A V NP003-HSRT C T R P S N N T R K S I I G P G Q T F Y NP128-MTCTN Q R A NP003- HSRA T G D I I G D I R Q A H C N I NP128-MTCT E.. K.....

NP003-HSRT C T R P S N N T R K S I I G P G Q T F Y NP041-IDUV. I.. N NP003-HSRA T G D I I G D I R Q A H C N I NP041-IDU Signature sequence pattern analysis 9 variable positions amino acid sequences between different risk groups All risk group had GPGQ pattern at v3 loop.