Current HIV Research - Volume 13, Issue 4, 2015

Background: Improved viral detections by the real time PCR over the manual assays have been reported by various manufacturers. However, discrepancies and discordance between different platforms targeting the same pathogen have also been observed at different settings. Methods: We used an analytical study design to compare the performance of the Cobas Taqman /Cobas Ampliprep version 2.0 against the standard Amplicor Monitor 1.5 using 200 routine clinical samples, in Abuja- Nigeria. Results: Taqman and Amplicor detected 118/200 (59%) and 83/200 (41.5%) samples respectively. Two of 83 samples (2.4%) undetectable by Cobas Taqman, were detectable by Roche Amplicor, while 5 of 37 samples (13.5%) which were undetectable by Amplicor using Taqman. Among the 81 detectable samples by both assays 4 samples (4.9%) had a log10 difference > 0.5 log copies, while 9 samples (11.1%) showed a wider discrepancy of >1 log10. Bland and Altman’s comparison shows no significant difference between the two methods (p=0.2825) and CI-0.06171 to 0.2087. Conclusion: We observed a remarkable improvement in the performance of COBAS AmpliPrep/COBAS TaqMan version 2.0 Assay over Amplicor Monitor version 1.5 in the quantification of HIV1 RNA viral load. Discrepancies of clinical significance, in the viral load between the two platforms were also recorded. The implications of the inability of the automated Taqman 2.0 to detect 2.4% of samples detectable by the Amplicor need to be considered by programs, clinicians and the manufacturers. Periodic evaluation of platforms to detect new circulating HIV subtypes within each locality is also recommended.

Background: Nigeria bears nearly 10% of the global burden of HIV/AIDS. Most of the AIDS patients dwell in the part of Nigeria known as the “North Central” geopolitical region. Sustaining HIV patients in this high risk region is critical for the overall success of the ART program in Nigeria. We assessed the level of adherence to ART and adherence determinants among participants who had been on ART for an average of three and half years. Methodology: Eligible study participants initiated HAART between 2004 and 2010. HAART regimens contained AZT/3TC +NVP or EFV; AZT/3TC/NVP; 3TC/NVP/d4T; TDF/FTC +EFV or NVP and TDF+3TC+LPV/r. A composite adherence measure defined as not missing a dose and taking the correct dose and adhering to the correct frequency and correct schedule of drug administration was used to assess self-reported adherence over a period of three days. Selfreported adherence was validated with viral load test. Base line adherence was fixed at ≥95% adherence level. Significant test was fixed at p<0.05. Results: We included 502 participants in the analysis. Median age for men was 42 years (IQR: 38 – 44 years) and women, 36 years (IQR: 30-40 years). Mean duration of therapy was 43 (16-70) months. Effective self-reported adherence was 97.3%. Only age and virologic suppression were significantly associated with adherence to ART. Forgetfullness (43%) was the major reason for non-adherence, while improvement in health condition (40%) was the main facilitator of adherence to the medications. Conclusion: Most participants achieved optimal adherence (≥95%) with high virologic suppression. Strategies to sustain optimal adherence, e.g., the use of fixed dose combinations (FDCs) and comprehensive adherence counselling should be maintained.

Background: Virological suppression is the main goal of antiretroviral therapy. To achieve this goal, efficient interventions that promote treatment adherence are needed. This study was aimed at exploring the impact of peer-education on virological outcomes in Northern Nigeria. Methods: A randomized controlled trial (RCT) among patients receiving antiretroviral treatment was conducted in 2 phases between August 2006 and January 2008 in the “largely Muslim” Northern Nigeria. Participants were randomized into one of three intervention arms: standard of care arm, a second arm which included daily reminders via alarm and follow-up calls from peer-educators, and adherence support by a home-based treatment partner; and a third arm which included second arm activities, plus home visits by peer-educators. We evaluated sociodemographic factors and adherence levels, measured using self-report and pharmacy (Rx) refill rates, as risk factors for viral load (VL) suppression. Results: Of the 600 participants (43% males), 276 were observed till the end of the study. There were no significant differences in mean log 10 VL between the intervention groups. At the end of entire follow-up period, 83% (229/276) who were not lost to follow-up achieved undetectable VL (< 400 copies/ml). In the multivariable analysis, age between 30-34 years (vs 18-24 years) and both baseline CD4 ranges between 100-199 cells/mm3 or 200-349 cells/mm3 (vs CD4 <100 cells/mm3) as positively associated with VL suppression while poor self-reported adherence and <95% Rx refill rates were negatively associated with VL suppression. Conclusion: High levels of viral suppression and low prevalence of drug resistance mutations (DRMs) were seen in this cohort participating in an ART adherence study in Northern Nigeria. Self-reported good adherence and optimal Rx refill rates were reported as significant predictors of VL suppression. Our findings indicate that ART adherence will improve significantly regardless of whether HIV-infected adults received peer-education-based medication adherence interventions or standard of care services.

Background: Despite improvement in HIV prevention of mother-to-child transmission (PMTCT), there are still over 1,500 African infants newly infected daily. PMTCT elimination requires antiretroviral therapy (ART) throughout pregnancy and breastfeeding periods, while early infant diagnosis (EID) of HIV implies early treatment for those infected. Our study aimed at assessing the utility of EID program data in evaluating the implementation of PMTCT program in Cameroon, and in identifying the efficacy of existing PMTCT interventions and breastfeeding options on the events of HIV vertical transmission. Methods: A study was conducted from 2010-2011 using PMTCT data from EID sites of six regions of Cameroon. PMTCT ARV regimens, breastfeeding options, and the child’s HIV DNA-polymerase chain reaction (PCR) results were recorded. Statistical analyses were performed using Mann Whitney U and Fisher exact tests, with p<0.05 considered significant. Results: A total of 2,505 mother-child pairs received ART, resulting is 4.3% (93) vertical transmission, against 31.3% (284/906) among mother-child pairs without exposure to any PMTCT intervention; p<0.00001. A statistically significant difference (p<0.00001) was also found between formula feeding (FF) (5.9%) versus exclusive breastfeeding (EBF) (12.5%), as well as between EBF versus mixed feeding (MF) (30%). With FF, when both mother-child pairs received PMTCT, only 2.9% (47/1603) vertical transmission was recorded versus 19.9% (48/241) for mother-child pairs without intervention; p<0.00001. Transmission rates were similar across infant age range [2.7% (10/376) for age ≤6 weeks, versus 2.5% (43/1807) for age >6 weeks-6 months]. Interestingly, babies aged 6 weeks receiving FF showed a significantly lower transmission rate (3.2%, 9/277) as compared to their counterparts with EBF (7.7%, 12/156); p<0.00001. Conclusion: Using EID dataset, it appears that considerable reduction in HIV MTCT may be achievable through access to ARV (option B+) and adequate infant feeding option (especially FF) in Cameroon. EID programme is therefore an effective routine approach for PMTCT programme evaluation in resource-limited settings.

Background: Hepatitis B (HBV) and Human Immunodeficiency virus (HIV) are both bloodborne viruses. Markers of either active or past HBV infection are present in many HIV infected patients. Worldwide, HBV prevalence varies geographically and endemicity is classified as low (<2%) or high (>8%). Genotypically, prevalence varies among different populations, with genotype A having a wide distribution. In Kenya, the prevalence of HIV-1/HBV co-infection ranges from 6-53% depending on the sub-population, with genotype A as the most common. Objective: To determine the prevalence and characterize HBV in HBV/HIV co-infected injecting drug users (IDUs) from Mombasa, Kenya. Methods: Blood samples were collected from HIV-infected IDUs in Mombasa, Kenya. Hepatitis B surface antigen (HBsAg) was tested by enzyme immunoassay (EIA). HBV DNA was extracted by SMITEST R&D kit. Polymerase chain reaction (PCR) was done; followed by population sequencing of HBV preS, core and full genome using specific primers. Analysis was done phylogenetically with reference sequences from the Genbank. Results: Seventy two HIV-positive samples were collected from IDUs in Mombasa in February and March 2010. Of these, 10 (13.89%) were HBsAg-positive by EIA. Nine of the 10 samples (12.5%) were PCR positive for HBV in the preS region; from these, four HBV full length sequences were obtained. Phylogenetic analysis showed that all belonged to genotype A1. Conclusion: The prevalence of HBV co-infection among HIV-infected IDUs in Mombasa, Kenya was 12.5%. Phylogenetically, sequences obtained from this study showed clusters that were distinct from reported Kenyan reference sequences from the Genbank. The findings point to an existence of a transmission network among IDUs in Mombasa. This further suggests that HBV genotypes in Kenya may be regionally diverse.

Background: Long-term retention is a crucial component of HIV care because treatment success can only be measured among retained patients. Understanding determinants of retention will inform retention strategies. We evaluated the correlates of retention in a large HIV program in Nigeria. Methods: We reviewed quality of care data for 5320 randomly selected HIV-positive adults aged ≥15 years enrolled in 37 treatment facilities in Nigeria between 2005 and 2009. Retention was described as having one or more clinic visits in the one year (2010) review period. Patient-related correlates of retention were determined using logistic regression. Results: 144 patients exited the program through deaths or transferrals. Of the 5176 with no documented exits, 3231 (62.4%) were retained (65.6% female; median age: 35.6 years). 2938 (75.8%) patients on ART, and 286 (23.4%) pre-ART patients were retained. Being on ART (OR=10.3, p<0.001), Age 30-60years (30 – 45 years: OR=1.36, p<0.001 and >45 – 60 years: OR=1.47, p<0.001) compared to patients <30 years; Female gender (OR=1.18, p=0.006), baseline CD4 cell count (100-350 cells/mm3: OR=1.24, p=0.006) vs <100cells/mm3 and lower WHO stage at baseline (WHO Stage IV, III, II: OR=0.50,0.51,0.77 respectively) vs Stage I were associated with retention. Among patients on ART, recent ART initiation 2008-09 (OR=1.73, p<0.001) vs 2005-07, being on ART for >6months (p<0.001) vs <6month and initiating ART on non-Stavudine based regimen (p<0.001) were also associated with retention. Conclusion: 3 out of 4 pre-ART patients and 1-in-4 ART patients were not retained in 37HIV treatment facilities in Nigeria. These findings provide insight that enables HIV programs integrate retention strategies at all stages of the HIV care continuum.

Background: Performance of Genotype MTBDRplus assay against Lowenstein Jensen (LJ) proportion method of Drug Susceptibility Testing (DST) in detection of resistance among clinical mycobacteria isolates to rifampicin (RMP) and isoniazid (INH) was evaluated in Ilorin, Nigeria. Methods: This retrospective study characterized one hundred mycobacteria isolates from pulmonary TB patients, stored on LJ medium and subcultured unto fresh LJ slants before being genotyped using MTBDRplus assay. DST was performed on the isolates against RMP, INH, Ethambutol and Streptomycin. Results: Genotype MTBDRplus identified 97% and 3% of the 100 isolates as Mycobacterium tuberculosis Complex (MTBC) and Non-Tuberculous Mycobacteria (NTM) respectively. Fourteen of the isolates, (14%) were resistant to RMP while 86% were sensitive by the genotypic method. Three of these 14 RMP-resistant isolates were NTMs while 11 were MTBC. Twelve (12%) of the 100 isolates were resistant to INH. Three INH-resistant isolates were NTMs, and 9 were MTBC. Phenotypically and genotypically, the 3 NTMs were resistant to RMP and INH and ten of the 97 MTBC strains were RMP-resistant. One RMP-phenotypically-sensitive strain was genotypically resistant to RMP. Six of the MTBC isolates were resistant to both RMP and INH by both methods. Most mutations occurred in the S-531L and S315T1 codons of rpoB and KatG genes of RMP and INH, respectively. Conclusion: The high specificity and positive predictive values recorded by MTBDRplus in our study make it suitable for use in the programmatic management of drug-resistant TB in resource-limited settings.

Background: The detection of acute HIV infection (AHI) among high risk populations can help reduce secondary transmission of HIV. The nucleic acid testing (NAT) can shorten the test window period by up to 7-12 days. In this study, we describe an in-house NAT based on the multiplex nested RT-PCR method to detect the HIV RNA. We also evaluated it in a high risk cohort in Beijing. Methods: Four primer pairs were designed and evaluated for the detection of different HIV-1 subtypes in group M. Multiplex RT-PCR and nested PCR were performed. The sensitivity, specialty, primers compatibility among HIV subtypes were evaluated simultaneously. In an MSM cohort in Beijing during a 3-year period, a total of 11,808 blood samples that were negative by ELISA or indeterminate by Western blot were analyzed by this multiplex nested RT-PCR with pooling strategy. Results: The multiplex nested RT-PCR was successfully applied for the detection of at least six HIV-1 subtypes. The sensitivity was 40 copies/ml and the specificity was 100%. A total of 29 people were tested HIV-1 positive with acute infection in a MSM cohort of Beijing during a 3 years period. Conclusion: This multiplex nested RT-PCR provides a useful tool for the rapid detection of acute HIV-1 infection. When used in combination with the 3rd generation ELISA, it can improve the detection rate of HIV infection, especially in the source limited regions.

Aichi viruses (AiV) have been detected in patients with diarrheal diseases (DD). The aim of this study was to assess AiV infection rates in hospitalized children with DD, including 123 HIV-1 seropositive and 125 HIV-1 seronegative patients, in two public pediatric hospitals in Rio de Janeiro, Brazil. AiV was investigated by nested RT-PCR. The AiV-positive samples were also tested for specie A rotavirus, norovirus, astrovirus, enteric adenovirus and bocavirus in order to assess co-infections. AiV parcial genome sequencing and phylogenetic analyses were performed. AiV were detected in 9/123 (7.32%) of the HIV-1 seropositive subjects and 1/125 (0.8%) of the HIV seronegative patients with DD (p = 0.019). The phylogenetic analysis of positive samples disclosed that: i) 13 samples were characterized as genotype A, with one of them being from the HIV-1 seronegative patient; ii) one sample from a HIV-1 seropositive patient was characterized as genotype B. AiV genotype A was grouped into 3 genetic clusters. Data suggest that AiV may be an opportunistic pathogen infecting children with AIDS and DD.