Predominant amino acid substitutions in NS5B gene of hepatitis C virus in blood donors and treatment-naïve hepatitis and HIV patients in Nigeria

Background: Hepatitis C virus (HCV) genome undergoes high rate of mutation, which results in generation of genetically diverse HCV isolates. There is paucity of data on mutations in the nonstructural 5b (NS5b) gene of circulating HCV and their implications in the Nigerian population. Here, we identified clinically-important mutations in HCV isolates, which may influence response to therapy and disease prognosis. Methodology: HCV RNA was extracted from a total of 301 blood samples collected from 99 symptomatic treatment-naïve hepatitis patients, 125 HIV-infected individuals and 77 asymptomatic blood donors in Ibadan, Nigeria. The RNA was reverse­transcribed to complimentary DNA and HCV NS5B gene amplified by nested PCR. The amplified products of 42 HCV were sequenced and sequences were aligned with those from GenBank and HCV databases in MEGA 7.0. Nucleotide sequences were translated to amino acids while substitutions in the amino acids were analyzed with reference to H77 prototype strain of HCV. Results: A total of 10 amino acid polymorphisms were observed from the 42 sequenced NS5B gene, with the major clinically-important amino acid mutations being S15G in 28 (66.7%) participants, T7N (24, 57.1%), G61R (23, 54.8%), S54L (22, 52.4%), G89E (14, 33.3%), T79M (12, 28.6%), and T711 (11, 26.2%). Others were Q67R (7, 16.7%), Q47H (7, 16.7%) and S84F (2, 4.8%). S15G/A/V mutations were more predominant in patients with HIV (76.9%, 10/13) followed by patients with clinical hepatitis (75.0%, 12/16) and blood donors (46.1%, 6/13). Q67R and T71I mutations were not predominant in patients with clinical hepatitis as they were detected in only 31.3% (5/16) and 43.8% (7/16) participants respectively, compared to S15G (75.0%, 12/16), S54L (68.8%, 11/16), G61R/E (68.8%, 11/16) and T7N/S (56.3%, 9/16). There was no statistically significant difference in the distribution of each of the 10 amino acid polymorphisms detected within patients with symptomatic clinical hepatitis (x 2=9.311, p=0.409), HIV-infected patients (x 2=13.431, p=0.1440) and asymptomatic blood donors (x 2=3.775, p=0.9256). Similarly, there was no significant difference in the distribution between the 3 categories of the study participants except for T79M mutation, which was significantly higher in HIV-infected patients (61.5%, 8/13) compared to patients with clinical hepatitis (18.8%, 3/16) and asymptomatic blood donors (7.7%, 1/13) (x 2=10.456, p=0.0054). Conclusion: Mutations in the NS5B gene could be associated with worse prognosis of the disease or antiviral failure due to viral resistance in patients undergoing therapy. The absence of Q47H mutations in majority of the study participants in our study implies that they will not respond well to daprevir and mericitabine. Screening of patients for pre-existing resistant mutations before commencement of therapy and monitoring during and after therapy are recommended.

Diverse genetic subtypes of HIV-1 among female sex workers in Ibadan, Nigeria

Genetic diversity is the hallmark of HIV-1 infection. It differs among geographical regions throughout the world. This study was undertaken to identify the predominant HIV-1 subtypes among infected female sex workers (FSWs) in Nigeria. Methods: Two hundred and fifty FSWs from brothels in Ibadan Nigeria were screened for HIV antibody using ELISA. All reactive samples were further tested by the Western Blot Techniques. Peripheral Blood Mononuclear Cells (PBMCs) were separated from the blood samples of each subject. Fragments of HIV Proviral DNA was amplified and genetic subtypes of HIV-1 was determined by direct sequencing of the env and gag genes of the viral genome followed by phylogenetic analysis . Results: The age of the FSWs ranged from 15 to 55 years old (Mean = 25.8years; SD =3.74). Majority were Nigerians while others (1.6 ) were from neighboring West Africa countries. Four ( 1.6 ) of the FSWs were active for less than one year as sex workers; and the mean length of sex work was 2.80 years ( Range = 1.0 - 15.0 years ). Sixty-four (25.6) of the 250 CSWs were positive for HIV-1 while 7 (2-8) had dual infections to HIV-1 / HIV-2. Among the 34 HIV-1 strains characterized by sequencing; 19 (55-9) were subtype G; 9 (26.5) CRF02_A/G; 3 (8.8) CRF06_cpx while 1 (2.9) each were identified as subtype C; CRF01_A/E and CRF09_cpx respectively. Nineteen (55.9) of the FSWs with subtype G had been active in the sex work for between one to five years. The youngest of the HIV -1 infected FSWs with sexual activity of less than a year had subtype G strain. There is a significant probability that infection with this subtype occurred with a short incubation period (p 0.05). Conclusion: This study showed a wide range of HIV- 1 subtypes among FSWs in Nigeria. The situation poses serious challenge for the design of HIV vaccine candidate for use in Nigeria

Service uptake and performance of the prevention of mother-to-child transmission (PMTCT) programme in Ibadan, Nigeria

The Prevention of Mother to Child Transmission (PMTCT) programme in the University College Hospital (UCH); Ibadan has been in existence for more than five years and has scaled up to other sites. The study evaluated the service uptake and performance of the programme using national key indicators. Antenatal and delivery records of women enrolled between July 2002 and June 2007 were reviewed. A total of 51952 women attended first antenatal visits and received HIV pre-test counselling. Of these; 51614 (99.5) accepted HIV test and 49134 (95.2) returned for their results. Out of the tested patients; 2152 (4.2) were identified to be HIV positive. Partners of positive patients accepting HIV testing were 361(16.7) with 87 (18.6) testing positive. There were a total of 942 deliveries out of which 39.2of the mothers and 95.2of the babies respectively received ARV prophylaxis. In all; 85.8(788/918) of the mothers opted for formula as the method of infant feeding. Out of the 303 babies eligible for ELISA testing; 68.3reported for the test and 17(8.7) tested positive. There has been progress in the programme; reflected in the increase in the number of new clients accessing the PMTCT service. However; partner testing and follow up of mother-infant pairs remain formidable challenges that deserve special attention

Genital ulcer diseases among HIV_infected female commercial sex workers in Ibadan, Nigeria

We evaluated the prevalence and association of Genital Ulcer Diseases (GUDs) among HIV-1 infected female commercial sex workers (FCSWs) in Ibadan; Nigeria. A total of 25O FCSWs from brothels in Ibadan were tested for presence of antibodies to HIV and Syphilis. Pelvic examinations for signs of sexually transmitted infections (STIs) were carried out on the subjects. Endocervical and high vaginal swabs were collected from each of the subjects to establish laboratory diagnosis of STIs. Their age ranged from 15 to 55 years (Mean = 25.8yrs; SD =3.74). Majority (246/250) were Nigerians; while 1.6were from neighboring West African countries. Sixty four (25.6) of the subjects were positive for HIV-1 while seven (2.8) had dual HIV-1/2 infection. Analysis of the STIs showed that 49 (19.6) of the CSWs had GUDs. Herpes genitalis was the commonest GUDs as it occurred in 25 (10) of the subjects. Other STIs identified were chancroid (5.6); syphilis (4.0) and lymphogranuloma venerum (LGV) (4). Sixteen (64.0) of the CSWs with herpes genitalis had HIV-1 infection. The risk ratio of herpes genitalis for HIV acquisition was 3.0 (95CI: 2.0 - 4.4). Syphilis and chancroid were also foundto be significantly associated with increased risk of HIV infection (p0.0001). The adjusted odd ratios for Herpes genitalis; chancroid; and syphilis were 3.7(1-13.0; p0.05); 19.8 (2.7-13 .0; p0.05) and 19.1(1-231.0; p 0.05) respectively. There is need to educate FCSWs continually to adopt safer sexual behaviours; seek early diagnosis and treatment of GUDs to reduce their risk of transmitting HIV infection

Occult HBV Infection among a Cohort of Nigerian Adults

Objective: To determine markers of HBV infection and detect the presence of its occult infection in serum of a cohort of adult Nigerians. Methodology: The study involved 28 adult Nigerians with viral hepatitis (Group 1) and 28 apparently healthy adult Nigerians as controls (Group 2). Their sera were assayed for HBsAg; HBeAg; anti-HBe; anti-HBc; anti-HBs; and anti-HCV; while HBV DNA was determined in 15 patients with chronic hepatitis. Significance of differences between the patients and control subjects was assessed using Chi-square test at a 95confidence level. Results: Sero-detection of HBsAg; HBeAg; anti-HBe and anti-HBc was higher among the patients compared to the controls. HBV infection was diagnosed by HBsAg (89) and a duo of HBsAg and anti-HBc (100) among the patients. Similarly; eleven and four types of different patterns of HBV markers were observed among the respective groups. Anti-HBe (9.5); anti-HBc (14.3); and anti-HBs (9.5) were detected among all the subjects who were sero-negative for HBsAg. HBV DNA was also detected in 86.7of the 15 patients with chronic hepatitis; while occult HBV infection was observed in 7.2of the patients and none (0) of the controls; p 0.05. Furthermore; HCV infection occurred among subjects with all the different patterns of HBV markers; except those with occult HBV infection and natural immunity to HBV. Conclusion: This study shows that occult HBV infection is present among Nigerian adults and determination of HBsAg; anti-HBc; anti-HBe; and HBV DNA will assist in its detection

Nigerian butchers and hepatitis B virus infection.

Various target groups have been identified in Nigeria for studying the prevalence of hepatitis B virus infection; however there is no information on its prevalence among workers in slaughter houses. This study determined the seroprevalence of hepatitis B virus infection in Nigerian butchers at Ibadan, and comprised 360 healthy Nigerian adult subjects (180 butchers, 180 traders as controls) selected by multistage stratified sampling. A questionnaire was used to collect relevant information and included points about risk behaviour. ELISA was used to detect the hepatitis B surface antigen in the serum; the seroprevalence rate in butchers and controls was 9.4% and 3.3%, respectively (p<0.05). Risk behaviour was seen more commonly in butchers than in controls. The presence of hepatitis B surface antigen in the serum was not related to the duration of occupational exposure or the number of partners. In summary, butchers comprise a high-risk occupational group for exposure to hepatitis B virus infection. We conclude that routine screening for parenterally acquired infections in this group is thus necessary in order to identify those who will require treatment and immunisation, especially against hepatitis B virus infection.

Mother-to-child transmission of different HIV-1 subtypes among ARV Naïve infected pregnant women in Nigeria

A taxa de transmissão materno-fetal (MTCT) do HIV bem como as implicações dos múltiplos subtipos circulantes para MTCT na Nigéria não são conhecidos. Este estudo foi realizado para determinar as diferentes taxas de MTCT dos subtipos de HIV-1 detectados entre gestantes infectadas antes que a administração da terapia ARV estivesse disponível na Nigéria. Vinte das mulheres HIV positivas que assinaram o consentimento durante a gravidez trouxeram seus filhos para seguimento na idade de 18-24 meses. Amostras de plasma de ambos, mãe e filho foram testadas para anticorpos HIV no Departamento de Virologia, UCH, Ibadan, Nigéria. Todas as amostras positivas (plasma e células mononucleares do sangue periférico - PBMCs) foram enviadas para o Instituto de Medicina Tropical da Antuérpia, Bélgica, onde os subtipos de vírus infectantes foram determinados utilizando-se a técnica HMA. No conjunto, uma taxa de transmissão de HIV, materno-fetal, de 45% foi encontrada neste grupo. Especificamente, 36,4%, 66,7% e 100% das mulheres infectadas com HIV-1 CRF02 (IbNg), G e B, respectivamente, transmitiram o vírus para seus filhos. Até onde pode ser verificado, este é o primeiro relato da taxa de MTCT do HIV na Nigéria. Os achados relatados neste trabalho serão uma útil referência para estimar a qualidade das terapêuticas atuais disponíveis para MTCT neste país.