1. Assessment of Genetic Diversity and drug resistance mutations to current classes of inhibitors circulating in HIV -1 infected individuals in Nigeria Negedu-momoh, O.R; Olonitola, O.S; Odama, L.E; Inabo, H.I; Mbah, H.A; Kasembeli, A.N; Inzaule, S.C; Odafe, S; Ojo, O.A; Khamofu, H. G; Agwale, S.M; Zeh, C ICASA 2011, Addis-Ababa

2. Introduction HIV prevalence in Nigeria is currently at 4.1% 3.1 million people are living with HIV/AIDS in 2010 with a total AIDS death of 215,130 Implemented WHO public health approach to ART delivery Studies have reported issues and complexity of use, genetic diversity and emergence of ARV resistance References 1.Federal Ministry of Health Technical Report (2010). National HIV/Syphilis Sentinel Survey among Pregnant Women Attending Ante-natal Clinics in Nigeria. 2. National Guidelines for HIV/AIDS Treatment and care in Adolescents and Adults. FMOH, Abuja-Nigeria. October 2010 3. Adeyi et al, 2006, AIDs in Nigeria. A nation on the Threshold

4. Introduction The predominant variants of HIV-1 in the Nigeria are subtype G and CRF02_AG Drug resistance (DR) mutations and polymorphic substitutions have been documented in treatment experienced and drug naïve patients CRF: Circulating Recombinant Form References 1. Abimiku et al. (1994). AIDS Res Hum Retroviruses, Nov 10 (11): 1581-3. 2. Agwale et al. (2001). Journal of Clin. Micro, June, p. 2110-2114. 3. Agwale et al. (2002). Vaccine 20: 2131-2139. 4. Agwale et al. (2006). AIDS Res Hum Retroviruses, volume 22, Number 1: pp 22-26. 5. Olaleye et al. (1993). J Infect Dis, 167: 710-714 6. Howard et al (1994). AIDS Res Hum Retroviruses, 10:1755-1757. 7. Idoko J, Njoku O. (2003). 13th International Conference on AIDS and STIs in Africa, Nairobi, Kenya. 8. Ojesina AI, (2006). AIDS Res Hum Retroviruses, 22: 770–779.

5. Objective To evaluate HIV-1 variants and drug resistance mutations in circulation to the current classes of antiretrovirals among HIV infected individuals in Nigeria

6. Methods A cross sectional study was conducted in 2007-2009 on 202 HIV infected patients attending ART clinics Studies covered 6 states including Federal Capital Territory in the 6 geopolitical zones in Nigeria Patients were randomly selected Patients were interviewed using questionnaire to collect some baseline data Ethical approval obtained from research ethics committee of Ahmadu Bello University (ABU) Zaria, Nigeria.

7. Methods (Cont’d) Viral RNA was quantified using b-DNA method (Versant) HIV-1 RNA V 3.0 Genotypic analysis was performed by Pol gene sequencing of 40 samples of a viral load>1000 copies/ ml In–house genotyping kits were used for the assay on ABI 3100 system at Centre for Disease Control/ Kenya Medical Research Institute (KEMRI) Phylogenetic analysis of the variants was done using Paup v 4.0 and subtyping tools (Rega HIV -1 v 2.0 &NCBI) References 1. Swofford DL. (2000). PAUP*, version 4.0b4, Sinauer, Sunderland, MA, 2. Rambaut A, Sequence alignment editor v 2.0 3. Ray S. Simplot for window v 2.5 4. McNulty et al., 2007; J. Clin. Microbiol. 45:517–521. 5.Yang C et al., 2010. J Clin Microbiol. 48(9): 3158-64 6. Johnson et al., 2009. Topics HIV Med. 17(5):138–145.

8. Patients ARV regimen

9. Results The median age -33 years, female patients - 130 (64%) Median plasma viral load - 66 copies/ml ART patients -160 (79%); drug naïve - 42 (21%) # on first line regimen- 153 (95%) Phylogenetic analysis revealed Subtypes: G (20; 50%), J (1; 2.5%) and CRF02_AG (14; 35%) CRF- 18-cpx (1, 2.5%), CRF06_cpx (3; 7.5%) and URF AD (1; 2.5%)

10. Results DR mutations to the RT inhibitors were observed in 7 individuals, 5 ART-experienced (5/15; 33%) and 2 drug naive (2/25; 8%) No major resistance mutations identified for PR inhibitors No multi-class resistance mutation to the 3 classes of ART drugs DR mutations observed for NRTIs were: M41L, L74I, V75M, F77L, Q151M, M184V, L210W, T215Y and for NNRTIs: K103N, Y181C, M230L, G190A/G Most commonly observed mutation were M184V (5/40;13%) and K103N (5/40;13%)

11. Results M41L and K103N resistant mutations were observed as NRTI and NNRTI in 2 drug naïve respectively Thymidine analogue mutations (TAMs): M41L, L210W,T215Y and Q151M mutation were observed in 3 ART patients with CRF02_AG and subtype G Polymorphisms were observed in the PR and the RT region of the pol gene Subtype specific polymorphisms A98AS /S/G (4/20; 20%;) and V82I (18/20;90%) were observed among subtype G strains

12. Summary of Drug Resistance mutations

13. Conclusions /Recommendation Diverse recombinant forms of HIV are on the increase URFs detected revealed a coinfection with diverse viral strains Transmission of DR virus among drug naïve is an important public health issue In view of the ART scale up, continuous assessment is needed to monitor evolving diverse strains, the resistance threshold and potential impact of drug resistance on ART programs in Nigeria

14. Thank you for listening

15. Acknowledgement CDC, HIV research Lab team, Kisumu, Kenya FMOH, Nigeria St Charles Borromeo, Onitsha; Muritala Mohammed Specialist Hospital, Kano; Mambilla Baptist Hospital, Gembu, Taraba; Daughters of Charity, Kubwa, Abuja (DOC); Federal Medical Center, Yola; Holy Family Catholic Hospital, Ikom; Mainland Hospital, Yaba and Maitama District Hospital, Abuja