Molecular Characterization of HIV-1 And Drug Resistance Among HIV-1 Infected Patients Attending Kayanza District Hospital, Burundi

This study was funded by the East Africa Public health Laboratory Network Project (EAPHLNP)/Burundi. Abstract Virological failure in management of HIV-1 infection has been reported to be between 11 to 24% after 12 months of treatment. Out of these, acquired or transmitted drug resistance mutations have been reported at 71% to 90% in Sub-Sahara Africa. In this cross-sectional study we aimed to determine virological failure and drug resistance mutations in HIV-1 infected patients on ART attending Kayanza district hospital, Burundi. Patients were recruited using a purposive sampling technique. After informed consent, 4mL of venous blood was collected from each patient. The blood was separated into plasma and cells for various laboratory assays. Plasma viral loads were quantified using the Abbott m2000rt system. Polymerase chain reaction using gene specific primers was done after extraction of nucleic acids from plasma with >1000 copies/ mL, followed by sequencing of all amplified samples. Drug resistance was determined using the IAS and Stanford University database, with phylogenetic analyses done using the neighbor joining method.Two hundred patients were recruited; 13% of the respondents had virological failure associated with multiple sex partners (adjusted odds ratio (aOR, 0.154 , p =0.016) and irregularity in taking medications (aOR: 0.4 , p=0.014). Fifteen samples were successfully sequenced; 80% (12/15) were HIV-1 subtype C, 7% (1/15) subtype A, and 13% (2/15) were HIV-1 subtype A1. Of these, 87.5% had at least one mutation (NRTI or NNRTI), while 12.5 % did not carry any Drug Resistance Mutations. The most common drug resistance mutations were M183V, T215V M41L, E44D, L74I, L210W and K65R, K103N, Y188H. The prevalence of virological failure was established at 13%.Our findings showed possible gaps in the last 90% of the 90-90-90 WHO target by 2020. The results highlight the need for intense viral load and resistance testing for patients to improve overall treatment outcome. Some strategies are needed to improve adherence counselling and drug resistance mutation testing should be implemented to monitor HIV-1 patients on ART in Burundi. Keywords: HIV-1, antiretroviral therapy, Virological failure, DRMs, Burundi. DOI: 10.7176/JBAH/9-10-05 Publication date:May 31st 201

Prevalence and Associated Risk Factors of Hepatitis B Virus Infections Among HIV-1 Infected Patients Attending the Comprehensive Care Clinic in Malindi Sub-County Hospital in Kenya

Human Immunodeficiency Virus (HIV) and Hepatitis B Virus (HBV) co-infections are common all over the world. Infection with HIV increases rates of HBV chronicity, prolong the time the HBV stays in circulation and increase liver-related morbidity. Factors such as intravenous drug use, multiple blood transfusions, presence of tattoos, unsafe sexual practices and being health workers have been implicated as drivers of infection & transmission of HBV & HIV. This study aimed to determine the prevalence and genotypes of HBV associated risk factors among HIV infected patients in a descriptive cross-sectional study. Malindi was chosen as a suitable study site because of the high numbers of residents involved in sex tourism as well as intravenous drug use. A structured questionnaire was used to capture social demographic data such as age, gender, employment status, occupation, the level of education and marital status, clinical history information such as duration since diagnosis with HIV, ART drug history, duration taking ARVs and baseline CD4 count and risk factors associated with HBV infections such as intravenous drug use, history of blood transfusion, tattooing/scarification, and the sexual history from 446 consenting randomly selected HIV infected participants. Five millilitres of whole blood was obtained from each participant, 50µl of which was used for CD4 cell counts using a flow cytometer. HBsAg serology was done using Diaspot® rapid diagnostic test and confirmed by Hepanostika® HBsAg Ultra ELISA kit (BioMérieux SA) and HBV DNA was extracted from all HBsAg positive samples. Nested polymerise chain (PCR) reaction and sequencing of the Pre S1 region was done. Sample sequences were compared with published HBV genotypes sequences from GenBank and Phylogenetic trees were constructed using the NJ Plot software using a PHB file created through DNA Database of Japan (DDBJ) to determine the HBV genotypes. Out of the 446 HIV positive participants, 126 (28.3%) were males and 320 (71.7%) females. Only 19/446 (4.26%) participants were positive for HBV based on rapid strip test while 22/446 (4.93%) participants had HBV based on ELISA. Twelve of the 22 ELISA positive samples were successfully amplified by PCR. Out of the 12 PCR positive samples 10 were successfully sequenced. Phylogenetic analysis revealed that 9/10 (90%) samples belonged to genotype A while 1/10 (10%) belonged to genotype E. Males (p=0.028) and intravenous drug use (p= 0.08) were significantly associated HBV infections. The high prevalence (4.9%) of HBV among HIV patients attending Malindi Sub-county hospital is most likely highly driven by intravenous drug use and multiple sexual partners among the male gender and is predominantly genotypes A and E which is similar to the general population. Keywords: Hepatitis B virus, HIV, Co-infection, HBsAg, genotypes, intravenous drug us

Chemokine Coreceptor-2 Gene Polymorphisms among HIV-1 Infected Individuals in Kenya

Chemokine Coreceptor-2 (CCR2) is an entry coreceptor for HIV-1. A mutation in the coding gene for this coreceptor, CCR2-64I, has been shown to be an important factor for delaying disease progression. In Kenya no studies have been done to determine the status of CCR2 gene polymorphisms among HIV-1 infected individuals. To determine the existence and distribution of CCR2 gene mutations and identify polymorphic groups of the coreceptor gene in the population, a cross-sectional study was conducted to analyze the differences in allelic frequencies of CCR2-64I among HIV-1 seropositive individuals. Blood samples were collected from HIV/AIDS screening centers and analyzed for the presence of CCR2-64I using restriction fragment length polymorphism (RFLP). One hundred and eighteen samples collected from different regions of the country were genotyped for the CCR2-64I mutation. Of these, 4 (3.4%) were homozygous mutants (I/I) and 21 (17.8%) were heterozygous (V/I). Ninety-three subjects (78.8%) were wild type (V/V). With the search for a preventive/therapeutic HIV vaccine elusive, the presence of CCR-2 gene polymorphisms that delay disease progression and prolong the lives of the infected in the Kenyan population may contribute to the growing evidence that host genetic factors are important in predicting susceptibility to HIV-1 infection