Diagnosis of dual human immunodeficiency virus types 1 and 2 infections in a resource-limited setting.

BACKGROUND: The presence of dual HIV-1/HIV-2 infection in Ghana and the different drug requirements for the treatment of HIV-1 and HIV-2 presents difficulties for the treatment of dual infections with both viruses. OBJECTIVES: To determine the prevalence of the dual sero-positive profile in treatment naive patients at a principal ART Clinic in Accra, Ghana and to investigate if rapid screening assays could be useful for diagnosis. DESIGN: A cross-sectional study. SETTING: A principal antiretroviral treatment centre in Accra, Ghana. SUBJECTS: Three hundred and twenty eight antiretroviral treatment naive patients. RESULTS: A total of 12 (3.7%) of patients seen were dual seropositive. There was a slight tendency of dual seropositive females being older than their HIV-1 counterparts (p = 0.088, CI = -10.833 to 0.753). Eight of the 12 of the dual seropositives were reactive for Genie II and were considered as possibly infected with both HIV-I and HIV-2. Seven (87.5%) of Genie II dual seropositives had strong intensities (> 1+) on both HIV-2 specific bands (sgp105 and gp36) on Innolia. CD4 counts were not significantly different in dual seropositives as compared to HIV-1 infected patients. CONCLUSIONS: Dual HIV-1/HIV-2 seropositives (and possibly infections) maybe common especially in older women. The Genie II will be useful as a supplemental rapid test for rapid and accurate differentiation of HIV-1 and HIV-2 antibodies at treatment centres.

Hepatitis E virus infection among pig handlers in Accra, Ghana.

OBJECTIVE: To determine the correlates of hepatitis E virus infection (HEV) in a sample of persons who work with pigs. DESIGN: Cross-sectional study. SETTING: Three pig farms in the Greater Accra Region of Ghana. SUBJECTS: Persons who work with pigs seen at the selected pig farms between the months of January and May 2008. RESULTS: One hundred and five persons who work with pigs voluntarily completed a risk-factor questionnaire and provided blood samples for unlinked anonymous testing for the presence of antibodies to HEV. The median age of participants was 36.5 +/- 15.0 years (range 12-65 years). Of the 105 subjects tested, HEV seroprevelance was 38.1%. On multivariate analysis, the independent determinants of HEV infection were being employed on the farm for less than six months (odds ratio (OR) 9.1; 95% confidence interval (95% CI) 1.0-81.4 and having piped water in the household and/or on the farm (OR 3.9; 95% CI 0.4-90.8). CONCLUSION: Consistent with similar studies worldwide, the results of our studies revealed a high prevalence of HEV infection in persons who work with pigs. Further studies need to be done to isolate, characterise the virus and define the clinical and epidemiological significance of HEV infection in this population.

Isolation and characterization of HIV-2 from an AIDS patient in Ghana.

This report describes the isolation and characterization of a retrovirus of the HIV-2 group from a Ghanaian AIDS patient which has different restriction patterns from previously reported HIV-2 viruses. The virus was morphologically very similar to HIV-1 and HIV-2, and had Mg2+-dependent reverse transcriptase. Like previous HIV isolates, it induced severe cytopathic effects in CD4-positive human lymphoid cell lines. Its major proteins were shown to be gp110, p66, p55, p41, gp32, p30 and p26 by Western blot analysis. In dot-blot hybridization experiments, the virus hybridized with a HIV-2 DNA probe, but not with HIV-1 and SIVagm probes in stringent conditions. These data indicate that this Ghanaian virus is a HIV-2 group virus. However, in a Southern blot hybridization experiment, the restriction patterns of this virus, designated HIV-2 [GH-1], were quite different from those of previously reported HIV-2 viruses from West Africa isolated at the Pasteur Institute.

Immunological reactivities of Ghanaian sera with HIV-1, HIV-2, and simian immunodeficiency virus SIVagm.

PIP: To learn more about the relative prevalence of viruses from the human immunodeficiency virus (HIV)/simian immunodeficiency virus (SIV) groups in Ghana, serum samples were collected in 1986 from 47 men and women with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC), 57 apparently healthy individuals, and 11 AIDS-free hospital inpatients. Western blot analysis revealed a total of 46 reactive sera. 43 of the 47 serum samples from those with AIDS or ARC were positive; 6 were seropositive for HIV-1, 18 for HIV-2, 17 for both HIV-1 and HIV-2, and the remaining 2 were not reactive with glycoproteins. Of the 2/10 individuals with mild symptoms of HIV infection who proved to be seropositive, 1 was positive for HIV-2 and 1 for HIV-1 and HIV-2. There were no seropositive reactions among the AIDS-free hospital patients, and only 1 such reaction (seropositive for HIV-2) among the healthy individuals. These findings indicate that both HIV-1 (6 cases in this series) and HIV-2 (20 cases) are responsible for the development of AIDS in Ghana, and that there is a high prevalence (18 cases) of cross-reactivity between the 2 viruses. There was no evidence of SIV infection. Further research is needed to determine whether these findings are a result of cross-reactivity between envelope proteins of HIV-1 and HIV-2, double infection of AIDS patients, or infection with a new variant strain. Since prostitutes comprised 25 of the 47 AIDS/ARC patients and 6 of the 10 with mild symptoms of HIV infection, they are an important target for preventive efforts.^ieng

Establishment of a phylogenetic survey system for AIDS-related lentiviruses and demonstration of a new HIV-2 subgroup.

We designed a universal primer (UNIPOL) for DNA amplification of AIDS-related viruses. The phylogenetic tree constructed from the presumed sequences amplified with UNIPOL was representative of the tree calculated from whole pol gene sequences so far reported. UNIPOL was able to amplify the sequences of all four major groups of primate lentiviruses and also that of a distinct virus from a Ghanaian patient with an AIDS-related complex, designated GH-2. This strain scarcely hybridizes with known HIV/simian immunodeficiency virus (SIV) DNA probes. Sequence analysis of the only amplified fragment revealed rapidly that GH-2 was quite similar to the recently reported HIV-2ALT(D205) and that these two viruses form a new subgroup distint from known HIV-2 and SIVmac/SIVsm in the large HIV-2 group. This system will be useful for further phylogenetic study of various primate lentiviruses.

Epidemic acute haemorrhagic conjunctivitis due to Coxsackie virus A24 variant in Ghana.

An outbreak of acute haemorrhagic conjunctivitis (AHC) occurred in Accra, Ghana, reaching a peak in July 1987. Individuals ranging from infants to adults over 50 years were infected, with those between 20 and 30 years being the most affected group. There was a female preponderance. Clinical features included conjunctivitis, subconjunctival haemorrhage and ocular pain. Some patients reported of blurred vision due to mild keratitis. Isolation of virus from clinical specimens of AHC patients was successful only in cells of human origin such as HeLa and FL. Coxsackie virus A24 variant (CA 24v) was identified as the aetiologic agent. This is the first report to associate CA 24v with an epidemic of AHC in Africa, south of the Sahara, which is outside the endemic area of Southeast Asia and the Caribbeans. This finding suggests that earlier outbreaks of AHC in Ghana and Africa may have been due to CA 24v but went undetected. The results of various tests performed during this study suggest that, at least, two antigenically different viruses of CA 24v circulated during the course of this epidemic.

Differentiation of dual seropositivity to HIV 1 and HIV 2 in Ghanaian sera using line immunoassay (INNOLIA).

Two hundred serum samples from AIDS-like patients, blood donors and miscellaneous patients were screened by ELISA for HIV 1-2. Repeatedly positive samples were confirmed using the Western Blot (WB). Twenty samples that were dual positive were further analysed by Innolia, which contains synthetic or recombinant antigens in HIV-1 and HIV-2. Only 5 of the samples were dual positive. The need for further analysis of dual WB results is re-emphasized.

PIP: A serologic study conducted in Ghana highlighted the importance of differentiating between HIV cross-reactivity and specific reactions directed against the virus itself. 200 serum samples collected from blood donors, patients suspected to have AIDS, and other patients were screened by enzyme-linked immunosorbent assay (ELISA) and, if reactive, confirmed by Western blot. The 20 samples that were dually positive for HIV-1 and HIV-2 according to Western blot were further analyzed by INNOLIA, which contains synthetic or recombinant antigens to HIV-1 and HIV-2. Only 5 of the 20 samples were dually positive. 10 samples were positive for HIV-1 alone, 4 were positive for HIV-2 alone, and 1 was indeterminate. The INNOLIA test offers a more specific test for differentiation than the Western blot, and it is simpler to use since results for both HIV-1 and HIV-2 are recorded on a single test strip.

Hepatitis B viral markers in Ghanaian pregnant women.

Seven hundred pregnant women referred for blood grouping were studied for the presence of some markers of Hepatitis B virus. After collection of personal data including place of permanent residence the sera were tested for the presence of HBs Ag and anti HBs. The sera positive for HBs Ag were tested for the "e" antigen and antibody. The results were analysed according to parity, age and geographical location. Six hundred and ninety-two results were available for analysis showing 464 (67.1%) to have antibodies. 215 (31.1%) had no marker whatsoever. 44 (6.4%) of the women were HBs Ag carriers. Eight of these had the e antigen indicating that 1.16 percent were therefore infective. Calculation was made to determine the extent of vertical transmission using the prevalence of the e antigen. 15% of HBV carrier rate in the population is due to perinatal transmission and this is significant. Parity, age and geographical location did not influence the prevalence of the various markers.

HIV-1 and HIV-2 in Ghana, west Africa: community surveys compared to surveys of pregnant women.

We conducted a population-based serosurvey of HIV-1/2 in 2,410 residents of two urban and two rural areas of southern Ghana, West Africa and compared the results to serosurveys of 1,417 pregnant women. Using conservative criteria, we found the prevalence of HIV-1/2 in community survey adults (> 15 years old) to be 1.5% in women and 1.0% in men. Among pregnant women, the prevalence was 9.3% in one rural area but only 1.0% in other areas. Many samples, especially among the pregnant women were HIV-1 and -2 dual reactive on serology. We attribute the disparity between prevalence in the community survey participants and pregnant women to local socio-economic factors. Women in this area have a tradition of working throughout West Africa as trader/ commercial sex workers. When pregnant or ill, they return to their home villages to be with their families. HIV surveillance programs which rely on pregnant women need to be sure that sampled populations are truly typical of the communities they are intended to represent.

PIP: HIV prevalence in urban and rural southern Ghana was investigated in a community-based serosurvey of 2410 people conducted in 1989-90 and an analysis of serum samples collected in 1987 from 1330 pregnant women attending antenatal clinics in the Eastern Region. None of the 906 children in the community-based survey was HIV-positive, including 6 born to HIV-infected women. Among the 1329 adults, 5 men (1.0%) and 12 women (1.5%) were infected. Among pregnant women, 100 (7.1%) were HIV-positive. At the four antenatal centers, HIV prevalence ranged from 0.6% to 10.3%. The high HIV prevalence recorded among pregnant women in the Manya-Krobo District (10.3%), a rural area away from major travel routes, likely reflects the tendency for ill women to return home to be cared for by their families. HIV surveillance programs that focus on pregnant women should be aware of this social phenomenon.

HIV seropositive cases presenting as urological patients at the Korle-Bu Teaching Hospital, Accra--a preliminary report.

Thirty-six patients (33 males, 3 females) were screened for HIV antibodies at the Urological clinics, at the Korle Bu Teaching Hospital, Accra because of chronic diarrhoea, unexplained weight loss, multiple sexual partners, recurrent sexually transmitted diseases, non resolution of symptoms or history of possible exposure to HIV. Ten patients (27.8%) were found to be HIV seropositive; 6 were confirmed as HIV-1, one as HIV-2 and 3 as both HIV-1 and HIV-2. The HIV seropositive patients presented as follows: recurrent cystitis and offensive vaginal discharge in 2 females, non-resolution or recurrence of urological symptoms and signs such as warts (genital, meatal and or urethral) in 4 males and recurrent penile ulcers in 4 males. This report indicates that physicians should be alert and screen high risk patients for both HIV-1 and HIV-2 antibodies.

The African AIDS case definition and HIV serology in medical in-patients at Komfo Anokye Teaching Hospital, Kumasi, Ghana.

In 914 consecutive medical admissions to Komfo Anokye Teaching Hospital, the prevalence of infection with Human Immunodeficiency Virus type I (HIV-I) and Human Immunodeficiency Virus type 2 (HIV-2) was 12.6%. The prevalence in females was twice that found in males. The infection rate was maximum in the age group 25-29 years for females (45%) and 30-34 years for males (29%). There were 7 cases infected with HIV-2 alone, 55 cases infected with HIV-I alone and 53 cases with dual infection. The cases with HIV-2 infection tended to be older than those with HIV-I infection. For detecting HIV seropositivity in our patients the World Health Organization recommended case definition for AIDS in Africa gave a sensitivity of 32%, a specificity of 93% and a positive predictive value of 42%. The case definition gave the highest specificity and positive predictive values when cases of tuberculosis were not included in the analysis.

PIP: During November 1989-January 1990 in Ghana, medical officers clinically examined and took blood samples from 914 consecutive admissions to Komfo Anokye Teaching Hospital in Kumasi to determine seroprevalence of HIV and different clinical features of HIV infection as well as to assess the value of the World Health Organization (WHO) clinical case definition for AIDS. 12.6% of the admissions were infected with HIV-1 and/or HIV-2. Females were more than two times likely to be infected with HIV than males (17.6% vs. 8.8%). Overall, 25-29 year old women had the highest HIV infection rate (45%). 30-34 year old men had the highest HIV infection rate among males. 56.5% of HIV-infected females and 30.4% of HIV-infected males were infected with both HIV-1 and HIV-2. 7 cases (5 females and 2 males) were infected with just HIV-2. Their ages ranged from 35 to 75 years. When the researchers applied the WHO clinical case definition to all HIV seropositive cases, they found its sensitivity to be 32%, specificity to be 93%, and positive predictive value to be 42%. Sputum-positive tuberculosis (TB) accounted for much of the false positives (28/53). 15% of the 76 sputum positive TB cases were HIV infected. When the researchers excluded all confirmed or suspected TB cases from the analysis, the specificity and positive predictive value increased to a maximum of 97% and 61%, respectively; sensitivity was 28%. Many HIV seropositive cases were not diagnosed with HIV infection either by the case definition or clinically by ward physicians. They probably were asymptomatic. Clinicians should suspect patients diagnosed with pneumonia or meningitis to be HIV infected, as was the case in this study (20% and 17% of HIV seropositive cases who were case definition negative, respectively).

Frequency of rubella antibodies in the population of some tropical African countries.

Data on the seroepidemiology of rubella in tropical Africa have been scarce. However, reports of serologic surveys in many countries within the last few years have changed this situation. Studies in Angola have shown an overall rate of positivity for rubella antibodies of 73% among females between the ages of 15 and 19 years. In the group between five and nine years of age, 74% had rubella antibodies. Therefore, Angolans acquired natural immunity before beginning school. In Ghana, greater than 70% of women of childbearing age had rubella antibodies, and in Nigeria 60% had antibodies. In The Gambia, by 14 years of age, 93% or more of the female population had immunity to rubella. Similar high levels were found in surveys conducted in Ethiopia, Upper Volta, and Uganda. However, a low level of 33% was recorded in Togo. The reports indicate that rubella infection is widespread and endemic on the African continent.

Seroepidemiological study of rotavirus infection in rural Ghana.

Children in a village in rural Ghana were found to have acquired antibody to rotavirus by the age of two years. The results of our study suggest that primary infection takes place early in infancy. Fifty-six per cent of the three to 20-months-old had a four-fold rise in antibody levels during the six months follow-up period and 72.2% of them had one or more episodes of diarrhoea compared with 42.8% of those whose antibody levels stayed the same or decreased. The major epidemic season is considered to be the dry season.

A study of the evolution of coxsackievirus A24 variant in Ghana by viral RNA fingerprinting analysis.

An epidemic of acute haemorrhagic conjunctivitis (AHC) caused by coxsackievirus A24 variant (CA24v) was reported in Accra, Ghana in May 1987. We studied 7 of the viral strains collected from May to November, 1987, by RNA genome fingerprinting. Pairwise comparisons of the oligonucleotide maps showed that genetic similarity among them ranged from between 60.0 to 84.7%. Using base sequence variations deduced from genetic similarity among the isolates, isolation time of the strains and the rate of nucleotide substitution (estimated in a previous paper, Miyamura et al., 1990), we calculated divergence times and constructed a phylogenetic tree. This tree indicated that all the 7 strains had diverged from each other from 11 to 26 months before the AHC epidemic in Accra. CA24v may have been introduced into the country or the neighbouring area, at least, more than two years earlier, i.e. in the early half of 1985.

Yellow fever in Ghana, 1977-80.

The 3-year yellow fever epidemic in Ghana that started in 1977 and tailed off in 1980 appears to be the heaviest on record. In all, 827 cases and 189 deaths were reported (a fatality rate of 22.8%), the patients coming from many villages scattered over 4 regions in the country. The distribution of cases and other epidemiological characteristics are described in this article.

Seasonality of rotavirus infection in Ghana.

Human rotavirus (HRV) infection and its seasonal distribution was studied over a 12-month period in Ghana. A total of 561 stool samples, 447 diarrhoea stools and 114 non-diarrhoea stools (controls), were obtained from children attending three polyclinics in Accra. Rotavirus was detected during 10 of the 12 months and showed a seasonal trend. It was high during the relatively cool dry months and low during the wet season. Peaks of infection were in February (26.2%) and September (24.5%). HRV was detected in 67 of 447 of the diarrhoea stools (15.0%) and in eight of 114 controls (7.0%). The HRV isolation rate was highest (20.2%) in the under-18-months age group. The RNA electropherotype of the HRV isolates was predominantly (83.6%) of the long type. Non-group A HRV was detected in 14.9% of the HRV-positive samples.

Isolation and characterization of a highly divergent HIV-2[GH-2]: generation of an infectious molecular clone and functional analysis of its rev-responsive element in response to primate retrovirus transactivators (Rev and Rex).

A highly divergent HIV-2 designated as HIV-2[GH-2] was obtained from an AIDS-related complex (ARC) patient in Ghana. A full-length molecular clone of this isolate was obtained and a biologically active clone was constructed. Its restriction pattern differed from that of prototype HIV-2[GH-1] in 25 of 35 restriction sites, but was strikingly similar to a previously characterized HIV-2 isolate from a Ghanaian (HIV-2ALT). The conserved integrase region (288-bp fragment) previously displayed 95% identity with that of ALT but 17-20% divergence from the HIV-2 prototype member, and a new distinct subgroup (HIV-2b) of HIV-2 consisting of GH-2 and ALT was postulated (Miura et al. 1991.) These isolates, however, were biologically distinguishable from each other by its replication capacity in a monocyte line, U937, in which GH-2 could not grow but ALT grew well. In addition, the nucleotide sequence of the LTR of this new isolate displays 21% divergence from that of prototype HIV-2[GH-1], but the core enhancer, Sp1 binding sites and TATA box were conserved. Although the 3' half of the env gene sequence which is deleted in HIV-2ALT clone showed 27% diversity from the prototype, functional differences in the rev-responsive element were not observed.

Human T-cell leukemia virus type I isolates from Gabon and Ghana: comparative analysis of proviral genomes.

Two isolates of human T-cell leukemia virus type I (HTLV-I) were obtained from lymphocyte cultures of a healthy carrier in Gabon and another in Ghana. Their proviruses were analyzed by Southern blot hybridization and compared with prototypical HTLV-I isolated in Japan and the United States. The provirus genomes of both strains were highly homologous to the prototype HTLV-I along the whole viral genome. The restriction endonuclease sites of the Ghanian isolate were almost identical with those of the prototype HTLV-I, but 10 of 26 sites of the Gabonese isolate were different from those of the prototype. Furthermore, the restriction map of the Gabonese isolate resembled those of a simian T-cell leukemia virus (STLV-I) isolated from a chimpanzee from Sierra Leone and a variant of HTLV-I from Zaire (HTLV-Ib) more closely than those of any other known HTLV-I. These results indicated the existence of some unique strains of HTLV-I transmitted among African people, and the importance of clarifying the origin and transmission of HTLV group viruses.

Seroepidemiologic studies of acute hemorrhagic conjunctivitis virus (enterovirus type 70) in West Africa. I. Studies with human sera from Ghana collected eight years after the first outbreak.

In 1969 and early 1970, during the pre-epidemic period of acute hemorrhagic conjunctivitis (AHC) in the upper north regions of Ghana, 191 human sera were collected. In 1977, eight years after the first epidemic of AHC in Ghana, 1008 human sera were also collected in various regions of Ghana. These sera were examined for virus neutralizing (VN) antibody against the J670/71 strain of enterovirus type 70 (EV70). In the pre-epidemic 1969-1970 sera, a low prevalence rate (6%) of VN antibody, with a titer of greater than or equal to 1:8, was found; the geometric mean titer (GMT) was also very low (1:1.85). In the post-epidemic 1977 sera, a high prevalence rate (53%) of VN antibody was found in the Ghanaian population, and the calculated GMT was also high (1:7.36). A higher antibody prevalence was detected in coastal (Accra: prevalence rate of 58% and GMT of (1:7.84), forest (Kumasi: 58% and GMT 1:7.52) and peri-forest (Tamale: 51% and GMT 1:7.46) areas than in the tropical savanna area (Bolgatanga: 41% and GMT 1:5.13). The demonstration of VN antibody in children under seven years of age showed that EV70 was still widely distributed and highly active in Ghana. The VN antibody found in human sera was predominantly IgG.