Historical epidemiology of hepatitis C virus in select countries-volume 4.

Due to the introduction of newer, more efficacious treatment options, there is a pressing need for policy makers and public health officials to develop or adapt national hepatitis C virus (HCV) control strategies to the changing epidemiological landscape. To do so, detailed, country-specific data are needed to characterize the burden of chronic HCV infection. In this study of 17 countries, a literature review of published and unpublished data on HCV prevalence, viraemia, genotype, age and gender distribution, liver transplants and diagnosis and treatment rates was conducted, and inputs were validated by expert consensus in each country. Viraemic prevalence in this study ranged from 0.2% in Hong Kong to 2.4% in Taiwan, while the largest viraemic populations were in Nigeria (2 597 000 cases) and Taiwan (569 000 cases). Diagnosis, treatment and liver transplant rates varied widely across the countries included in this analysis, as did the availability of reliable data. Addressing data gaps will be critical for the development of future strategies to manage and minimize the disease burden of hepatitis C.

Strategies to manage hepatitis C virus infection disease burden-Volume 4.

The hepatitis C virus (HCV) epidemic was forecasted through 2030 for 17 countries in Africa, Asia, Europe, Latin America and the Middle East, and interventions for achieving the Global Health Sector Strategy on viral hepatitis targets-"WHO Targets" (65% reduction in HCV-related deaths, 90% reduction in new infections and 90% of infections diagnosed by 2030) were considered. Scaling up treatment and diagnosis rates over time would be required to achieve these targets in all but one country, even with the introduction of high SVR therapies. The scenarios developed to achieve the WHO Targets in all countries studied assumed the implementation of national policies to prevent new infections and to diagnose current infections through screening.

The present and future disease burden of hepatitis C virus infections with today's treatment paradigm: Volume 4.

Factors influencing the morbidity and mortality associated with viremic hepatitis C virus (HCV) infection change over time and place, making it difficult to compare reported estimates. Models were developed for 17 countries (Bahrain, Bulgaria, Cameroon, Colombia, Croatia, Dominican Republic, Ethiopia, Ghana, Hong Kong, Jordan, Kazakhstan, Malaysia, Morocco, Nigeria, Qatar and Taiwan) to quantify and characterize the viremic population as well as forecast the changes in the infected population and the corresponding disease burden from 2015 to 2030. Model inputs were agreed upon through expert consensus, and a standardized methodology was followed to allow for comparison across countries. The viremic prevalence is expected to remain constant or decline in all but four countries (Ethiopia, Ghana, Jordan and Oman); however, HCV-related morbidity and mortality will increase in all countries except Qatar and Taiwan. In Qatar, the high-treatment rate will contribute to a reduction in total cases and HCV-related morbidity by 2030. In the remaining countries, however, the current treatment paradigm will be insufficient to achieve large reductions in HCV-related morbidity and mortality.

Prevalence of enteric infections among hospitalized patients in two referral hospitals in Ghana.

BACKGROUND: Diarrhea is an important cause of morbidity and mortality worldwide. In Africa and Ghana in particular, it is estimated to contribute directly to 19 and 25% of pediatric mortality among children under 5 years, respectively. METHODS: Surveillance for hospitalized acute diarrheal illness was initiated in November 2010 through October 2012 in a referral hospital in southern Ghana, and a teaching hospital in northern Ghana. Consenting hospitalized patients who met a standardized case definition for acute diarrheal illness provided demographic and epidemiologic data. Stool samples were collected and tested by culture for bacteria and by enzyme immunoassays for a panel of viruses and parasites. RESULTS: A total of 429 patients were enrolled; 216 (50.3%) were under 5 years, and 221 (51.5%) were females. Stool samples were received from 153 patients. Culture isolates included Shigella sp., Salmonella spp., Plesiomonas sp. and Vibrio cholerae. Of 147 samples tested for viruses, 41 (27.9%) were positive for rotaviruses, 11 (7.5%) for astroviruses, 10 (6.8%) for noroviruses, and 8 (5.4%) for adenoviruses. Of 116 samples tested for parasitic infections; 4 (3.4%) were positive for Cryptosporidium sp. and 3 (2.6%) for Giardia lamblia. Of the enrolled patients, 78.8% had taken antibiotics prior to sample collection. CONCLUSIONS: Diarrheal pathogens were identified across all ages, however, predominantly (81%) in the children under 5 years of age. This study also detected high antibiotic use which has the potential of increasing antibiotic resistance. The most common enteric pathogen detected (49.4%) was rotavirus.

Plasma concentrations of transforming growth factor beta 1 in non-progressive HIV-1 infection correlates with markers of disease progression.

The human immunodeficiency virus (HIV) infection shows variable rate of disease progression. The underlying biological and molecular mechanisms involved in determining progression of HIV infection are not fully understood. The aims of this study were to determine plasma concentrations of active TGF ß 1, Th1 and Th2 cytokines in patients with non-progressive and those with progressive HIV-1 infection, as well as to determine if there is an association of these cytokines to disease progression. In a cross-sectional study of 61 HIV-1 infected individuals categorized according to disease progression as having non-progressive HIV-1 infection (n=14) and progressive infection (n=47), plasma levels of active TGF ß 1, INF-γ, TNF-α, IL-10, IL-1ß, IL-12p70 and IL-13 were compared with HIV uninfected healthy controls (n=12). Plasma concentration of these cytokines was measured using a highly sensitive luminex200 XMAP assay. Pearson correlation test was used to assess the correlation of cytokines with CD4+ and CD8+ T cells, CD4:CD8 ratio and plasma HIV-1 RNA in the different study groups. Plasma concentrations of TGF ß 1 and IL-10 were significantly decreased while IL-1ß, IL-12p70 and TNF-α were increased in patients with non-progressive HIV-1 infection compared to patients with progressive infection. Plasma levels of TGF ß 1 and IL-10 showed an inverse correlation with CD8+ T cell counts and CD4:CD8 ratios in patients with non-progressive HIV-1 infection, while plasma HIV-1 RNA positively correlated with CD4+ T cell counts. Plasma levels of TNF-α, IL-1ß, IL-12p70 and IL-13 positively correlated with CD4+ T cell counts and inversely correlated with plasma HIV-1 RNA, CD8+ T cell count and CD4:CD8 ratio in patients with non-progressive infection. The correlation of cytokines to the state of T-lymphocyte and plasma HIV-1 RNA found in this study may provide insight into the role of cytokines in both progressive and non-progressive HIV-1 infection. Additionally, these findings may have implications for systemic cytokine-based therapies in HIV-1 infection.

THE EVALUATION OF DOMESTIC DUCKS AS POTENTIAL RESERVOIR OF AVIAN INFLUENZA VIRUS IN POST HPAI H5N1 OUTBREAK AREA, SUNYANI MUNICIPALITY, BRONG AHAFO REGION OF GHANA.

BACKGROUND: Avian influenza (AI) is an important zoonotic disease responsible for significant losses in most sub-Saharan countries. However, the role of poultry other than chicken in the epidemiology of the disease, especially after the first AI outbreak in Ghana, has not been fully elucidated. The obiective of this study is to determine whether the AI virus infection that was reported in the area in May 2007 was circulating silently in ducks in nine randomly selected farms in the Sunyani Municipality, Ghana. MATERIALS AND METHODS: The sample size was calculated using Epi info version 3.4.1 at 95% confidence level, absolute precision of 5% and assuming 0.5 prevalence of Avian Influenza A virus in ducks. Samples collection was done simultaneously with questionnaire administration to farmers. A total of 526 samples made up of 384 cloacal swabs and 142 feather tissues from ducks from a commercial duck farm, seven backyard holdings and one live birds market in six randomly selected communities in the Sunyani Municipality, Brong Ahafo region of Ghana. The samples were processed and subjected to Influenza Type A Matrix Gene analysis using RRT-PCR. RESULTS: All the 526 samples subiected to Influenza Type A Matrix Gene analysis using RRT-PCR were negative for Influenza Type A viruses. However, it was observed that bio-security practices which are keys to reintroduction of the virus in the area were not adhered to in 89 % of the sites investigated. Our finding also revealed that only the commercial farm investigated in this study complied with fifteen (78.9%) of the nineteen different farm practices observed. CONCLUSION: Though AI was not detected in the ducks sampled, there is the need for continuous surveillance and education of stakeholders on standard bio-security and farm management practices in the area.

CD4+ T cell counts in initiation of antiretroviral therapy in HIV infected asymptomatic individuals; controversies and inconsistencies.

The primary goal when devising strategies to define the start of therapy in HIV infected individuals is to avoid HIV disease progression and toxicity from antiretroviral therapy (ART). Intermediate goals includes, avoiding resistance by suppressing HIV replication, reducing transmission, limiting spread and diversity of HIV within the body and protecting the immune system from harm. The question of how early or late to start ART and achieve both primary and intermediate goals has dominated HIV research. The distinction between early and late treatment of HIV infection is currently a matter of CD4+ T cells count, a marker of immune status, rather than on viral load, a marker of virus replication. Discussions about respective benefits of early or delayed therapy, as well as the best CD4+ T cell threshold during the course of HIV infection at which ART is initiated remains inconclusive. Guidelines issued by various agencies, provide different initiation recommendations. This can be confusing for clinicians and policy-makers when determining the best time to initiate therapy. Optimizing ART initiation strategies are clearly complex and must be balanced between individual and broader public health needs. This review assesses available data that contributes to the debate on optimal time to initiate therapy in HIV-infected asymptomatic individuals. We also review reports on CD4+ T cell threshold to guide initiation of ART and finally discuss arguments for and against early or late initiation of ART.

Low level of transmitted HIV Drug resistance at two HIV care centres in Ghana: a threshold survey.

BACKGROUND: As access to antiretroviral therapy (ART) increases, the emergence and transmission of HIV drug resistant strains becomes a major problem. The World Health Organization (WHO) therefore recommends an initial minimum-resource method to signal when transmitted HIV drug resistance (HIVDR) requires action. OBJECTIVE: This survey sought to generate information on the presence of HIV drug-resistant strains in the locality where Ghana's ART for HIV was first introduced. METHODS: The Ghana HIVDR threshold survey (TS) was conducted and analyzed according to WHO strategy for surveillance of HIVDR in the Eastern Region of Ghana. Sixty (60) plasma specimens were collected from 2007 to 2009 by an unlinked anonymous method from HIV seropositive pregnant women, aged between 15 to24 years, who were with their first pregnancy and ART naive. Genotyping was done as follows; Ribonucleic acid (RNA) was extracted from the samples and the protease (PR) and reverse transcriptase (RT) genes amplified and sequenced. The sequences were then analyzed for HIV drug resistance mutations using Stanford University HIV Drug Resistance Database. RESULTS: Only two individuals were found with major HIVDR mutations: one each in the PR and RT genes. Thus the level of HIVDR in the study population in 2009 was classified as low (< 5%). CONCLUSION: As at February 2009, transmitted drug resistance was not a serious problem in the Eastern Region of Ghana. However, it is important to continue monitoring tHIVDR in order to understand the dynamics of the evolution of HIV drug resistance in the country.

The first cases of Lassa fever in Ghana.

Lassa fever is a zoonotic disease endemic in West Africa but with no previous case reported in Ghana. We describe the first two laboratory confirmed cases of Lassa fever from the Ashanti Region of Ghana detected in October and December, 2011.

Infectious DNA clone of HIV type 1 A/G recombinant (CRF02_AG) replicable in peripheral blood mononuclear cells.

We constructed an infectious DNA clone of the HIV-1 A/G recombinant 97GH-AG2, which was isolated in Ghana in 1997 and was classified originally as subtype A. By phylogenetic and recombination breakpoint analyses, p97GH-AG2 was grouped in the circulating form of A/G recombinants (CRF02_AG) and was found to contain the least amount of subtype G-derived region among the known CRF02_AG HIV-1 DNAs. This result suggests that CRF02_AG may be a predominant form in Ghana. Virions produced by transfection of p97GH-AG2 into 293T cells grew in phytohemagglutinin-stimulated peripheral blood mononuclear cells (PBMCs). 97GH-AG2 also replicated efficiently in CCR5-expressing HeLa cells, MAGIC5, but only weakly in the parent MAGI cells, indicating that 97GH-AG2 uses mostly CCR5 as a coreceptor. Isolation of the first HIV-1 (CRF02_AG) DNA clone that replicates in PBMCs will accelerate the molecular analysis of this subtype.

Suitability of a rapid immunochromatographic test for detection of antibodies to human immunodeficiency virus in Ghana, West Africa.

In West African countries such as Ghana, efficient human immunodeficiency virus (HIV) testing is a priority in the fight against AIDS. A new immunochromatographic rapid test, Determine HIV-1/2 (Abbott Diagnostics, North Chicago, Ill.), that detects antibodies against HIV type 1 (HIV-1) and/or HIV-2 was evaluated using Ghanaian blood samples. Two hundred four serum and/or plasma specimens were tested. HIV screening was done by a particle agglutination test and confirmed by a Western blot (WB) test as the "gold standard." The results revealed 125 HIV-seropositive AIDS patients, 75 HIV-seronegative healthy individuals, and 4 individuals for whom the HIV-1 result was indeterminate. The results obtained by the Determine HIV-1/2 assay and Diagnostic HIV SPOT (Genelabs), which is currently widely used in many districts in Ghana, were compared with those of the WB test, excluding the four HIV-1-indeterminate samples. The sensitivity of the Determine HIV-1/2 assay was 100%, compared with 98.0% for the HIV SPOT assay. The specificity was 100% for both tests. Determine HIV-1/2 is a single-step assay and was found to be rapid and easy to perform without any special equipment. It was highly sensitive and specific. The kit can be applied without electricity and water supplies, making it suitable for the detection of HIV antibodies especially in the rural areas of Ghana, West Africa.

Isolation and characterization of a full-length molecular DNA clone of Ghanaian HIV type 1 intersubtype A/G recombinant CRF02_AG, which is replication competent in a restricted host range.

We have isolated a replication-competent, full-length molecular clone of HIV-1 CRF02_AG, designated p97GH-AG1, by reconstituting two separately amplified genomic regions of an HIV-1 provirus of a 1997 Ghanaian isolate. The phylogenetic and recombination breakpoint analyses revealed that 97GH-AG1 had an A/G recombinant structure similar to that of prototype Nigerian isolate IbNG. The 17-nucleotide insertion downstream of the primer-binding site appeared to be a common sequence signature specific to most CRF02_AG strains, including 97GH-AG1. 97GH-AG1 showed an R5 phenotype and exerted productive infection in both HOS and NP2 cell infectivity assays, whereas it failed to show a detectable level of progeny production in peripheral blood mononuclear cells (PBMCs). The data may suggest the presence of unknown determinant(s) that dictate efficient replication in PBMCs, but that are not required for replication in immortalized cell lines.

Genetic analysis of HIV type 2 from Ghana and Guinea-Bissau, West Africa.

The phylogenetic variability of part of the long terminal repeat (LTR) region of HIV-2 strains isolated in 1995 from five individuals residing in Bissau, the capital city of Guinea-Bissau, and collected from seven persons from Kumasi, Ghana in 1996-1997, was analyzed. All Guinean samples and all but one Ghanaian sample clustered with HIV-2 subtype A. One Ghanaian sample (14%) was classified as HIV-2 subtype B. This study adds to previous reports on HIV-2 subtype distribution in West Africa indicating local prevalence of HIV-2 subtype B in Ivory Coast and neighboring Ghana.

Relationship between immunoclinical status and prevalence of viral sexually transmitted diseases among human immunodeficiency virus-1 seropositive patients in Ghana.

In view of the strong association between the acquired immunodeficiency syndrome (AIDS) and sexually transmitted diseases (STDs), we screened 182 human immunodeficiency virus (HIV)-1 infected patients over a 15-month period for serological markers to previously encountered or current STDs, most of viral etiology. The relationship between their immunological and clinical status and the prevalence of STDs was assessed and compared with that of 88 HIV-seronegative patients. Hepatitis B virus and Treponema pallidum were the most frequently occurring pathogens in both HIV-1-infected and HIV-seronegative patients. Hepatitis C virus (HCV) infection was also observed in both groups, but no HIV-seronegative patient was infected with human T-lymphotropic virus type 1 (HTLV-1). The Centers for Disease Control clinical staging of A1 through C3, representing asymptomatic to severe AIDS conditions, was observed in HIV-1 patients with or without STDs. A mean CD4 count of 288 cells per microliter (95% CI of 237-340 cells per microliter) in HIV-1 patients was significantly lower (P < 0.05) than that in HIV-seronegative individuals with 1019 cells per microliter (95% CI of 924-1115 cells per microliter), irrespective of whether subjects in either group had previous or current STDs. The mean CD4 count of patients with a single infection from HIV-1 was not significantly different (P = 0.36) from that of HIV-1 patients with multiple infections. HIV-1 infection alone appears to be responsible for the marked immunodeficiency status of seropositive patients observed in this study.

Seroreactivity clarification and viral load quantitation in HIV-1 and HIV-2 infections in Ghana.

In Ghana, West Africa, the prevalence of dual HIV-1 and HIV-2 infections remains to be clarified, and HIV viral load measurement is yet to be established. Conventional assays for HIV-1 RNA measurements have been limited specifically to HIV-1 subtype B, preventing their utilization for Ghana where HIV-1 subtypes A, D and G are prevalent. Therefore, we set out to distinguish the types of HIV infection existing in Ghana so as to determine the extent of actual dual infections, and to measure plasma HIV-1 RNA. Blood samples were collected from 563 sick and healthy Ghanaians who visited hospitals in 1996 and 1997. After T cells were counted, HIV antibody was screened and confirmed by six different commercial assays and one in-house test. Nested PCR was then used to verify HIV-1 and HIV-2 presence by type-specific primers. Plasma HIV-1 RNA was measured by an improved commercial RT-PCR assay, sensitive to all HIV-1 group M subtypes. HIV-1 alone (89%) clearly dominated over HIV-2 alone (2%), and HIV-1 and HIV-2 dual infections were found in 9%. Valid viral load measurements were obtained on test plasma representing the main HIV-1 subtype (A) prevailing in Ghana. A high amount of HIV-1 RNA (5.9 mean log10 RNA copies/ml) was observed in the typical stages of HIV infection represented by groups of CD44 cell counts. We have clarified the seroprevalence of HIV-1 and HIV-2 amongst HIV seropositives, and the high viral load of HIV-1 reflects its influence on AIDS in Ghana.

High-level production of alternatively spliced soluble interleukin-6 receptor in serum of patients with adult T-cell leukaemia/HTLV-I-associated myelopathy.

We have previously shown, using human T-cell lymphocytotrophic virus-I (HTLV-I)-infected cell lines, that soluble interleukin-6 receptor (sIL-6R) is generated through an alternative splicing mechanism. In this study, we examined human sera for the presence of alternatively spliced soluble IL-6R (AS-sIL-6R). We produced a monoclonal antibody (mAb) recognizing the unique sequence of AS-sIL-6R peptide, generated by an altered reading frame. We also made recombinant AS-sIL-6R protein in Spodoptera frugiperda-9 (Sf-9) cells carrying baculovirus, which encoded altered sIL-6R or conventional IL-6R cDNA. mAbs specifically recognized AS-sIL-6R, but not conventional IL-6R, as demonstrated by Western blot analyses, fluorescence-activated cell sorter, immunofluorescence analyses and enzyme-linked immunosorbent assay (ELISA). We adapted an ELISA system and used it for detection of altered sIL-6R in sera from 23 healthy persons, 12 patients with adult T-cell leukaemia (ATL) and 13 patients with HTLV-I-associated myelopathy (HAM). Serum levels of AS-sIL-6R were 6.4 or 6.1 times greater in ATL (28.7+/-20.4 ng/ml, P<0.0001) and in HAM patients (27.5+/-12.1 ng/ml, P<0.0001) than in healthy individuals (4.5+/-2.1 ng/ml). High levels of AS-sIL-6R were also observed in plasma from rheumatoid arthritis patients and in persons with elevated levels of alanine aminotransferase (ALT), antinuclear antibody (ANA), or alpha-fetoprotein (AFP). However, in human immunodeficiency virus-1 (HIV-1), hepatitis B virus (HBV) or hepatitis C virus (HCV)-infected individuals, AS-sIL-6R levels were not elevated. In this study, we confirmed that AS-sIL-6R is indeed present in human sera. These observations suggest that alternative splicing of IL-6R mRNA is of consequence in ATL, HAM and in some autoimmune diseases. The HTLV-I-infected T cells appeared to play an important role in AS-sIL-6R production.

Phylogenetic analysis of human immunodeficiency virus 1 in Ghana.

Eleven human immunodeficiency virus 1 (HIV-1) isolates from Ghanaian acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) patients obtained by our serosurvey in 1986-1994 were genomically analyzed and phylogenetically compared with other known strains. A phylogenetic tree constructed by analyzing the env region indicated that heterogeneous HIV-1 strains were circulating in Ghana and the majority of them (9 of 11 isolates) belonged to clade (subtype) A which is now furiously epidemic in Africa. Another isolate (1 of 11) belonged to clade D, and the remaining one (1 of 11) belonged to "clade G". This "clade G" virus grouped by the env analysis belonged to clade A by its pol sequence, suggesting an A/G intersubtype recombinant. The characteristic sequences in the V3 tip which have not yet been reported were observed in these Ghanaian isolates, which should be taken into account for future vaccine programs.

PIP: The molecular epidemiology of HIV-1 in Ghana was investigated through genomic and phylogenetic analysis of isolates from 11 AIDS or AIDS-related complex patients obtained in 1986-94. A phylogenetic tree constructed by analyzing the env region indicated that heterogeneous HIV-1 strains are circulating in Ghana. 9 of the isolates belonged to clade A, 1 to subtype D, and 1 to "clade G"--an A/G intersubtype recombinant. The V3 loops of all isolates were composed of 35 amino acid residues--a characteristic not previously described. These molecular data on the genetic variability of the envelope glycoprotein of HIV-1 should be useful for future vaccine studies in West Africa.