Implementation of an Africa-specific donor health questionnaire for human immunodeficiency virus risk screening.

BACKGROUND AND OBJECTIVES: We had previously developed an Africa-specific donor health questionnaire (ASDHQ) based on local risk factors and designed a scoring scheme. This study assessed the performance of a new donor health questionnaire by comparing the human immunodeficiency virus (HIV) status in accepted versus deferred donors by ASDHQ and comparing the rate of risk deferrals with historical data. MATERIALS AND METHODS: Data were collected during a cross-sectional study conducted over 15 months at three referral-hospital-based blood services in Cameroon. ASDHQ was administered to blood donors aged 18-65 years in the same screening conditions as the routine questionnaire. The main outcomes of the study were ASDHQ sensitivity and specificity with regard to HIV laboratory testing as well as donor deferral rates for each of the routine screening algorithms and for ASDHQ. RESULTS: Overall, 71/11,120 (0.6%) were confirmed as HIV positive. The mean ASDHQ score was 95.80 ± 4.4 in HIV-negative donors and 94.80 ± 4.4 in HIV-positive donors (p = 0.05). The optimal cut-off provided by the receiver operating characteristic (ROC) curve for the best performance of ASDHQ was 95.04. Using this optimal cut-off, the ASDHQ sensitivity and specificity were 57% and 53%, respectively (area under curve = 0.58 [0.51, 0.64], p = 0.028). Using ASDHQ, the HIV prevalence was 0.7% in deferred donors and 0.6% in accepted donors. CONCLUSION: ASDHQ might be efficient only in specific conditions that maximize truthful donor responses, requiring each blood service to create an environment of trust and transparency to increase donor compliance and improve the accuracy of the questionnaire.

Transfusion management of severe anaemia in African children: a consensus algorithm.

The phase III Transfusion and Treatment of severe anaemia in African Children Trial (TRACT) found that conservative management of uncomplicated severe anaemia [haemoglobin (Hb) 40-60 g/l] was safe, and that transfusion volume (20 vs. 30 ml/kg whole blood equivalent) for children with severe anaemia (Hb <60 g/l) had strong but opposing effects on mortality, depending on fever status (>37·5°C). In 2020 a stakeholder meeting of paediatric and blood transfusion groups from Africa reviewed the results and additional analyses. Among all 3196 children receiving an initial transfusion there was no evidence that nutritional status, presence of shock, malaria parasite burden or sickle cell disease status influenced outcomes or modified the interaction with fever status on volume required. Fever status at the time of ordering blood was a reliable determinant of volume required for optimal outcome. Elevated heart and respiratory rates normalised irrespective of transfusion volume and without diuretics. By consensus, a transfusion management algorithm was developed, incorporating three additional measurements of Hb post-admission, alongside clinical monitoring. The proposed algorithm should help clinicians safely implement findings from TRACT. Further research should assess its implementation in routine clinical practice.

Trends in blood donations, blood donors' knowledge, practices and expectations during the COVID-19 pandemic in Cameroon.

INTRODUCTION: The COVID-19 pandemic, caused by a novel coronavirus, has already affected over 99 062 people in 53 African countries and killed 3082. The pandemic threatens blood supply but we do not yet know its impact on blood donations or on the perceptions and expectations of donors. METHODS: We conducted a cross-sectional study in four hospital-based blood services in Cameroon, using a survey design and focusing on the subjective and cultural aspects of donors. Using a semi-structured questionnaire, we collected the participants' responses as to their understanding of COVID-19 and of current protection measures, and their expectations. Data on trends of blood donations were collected retrospectively for the period from 1st January to 30th April 2019 and the same period in 2020. RESULTS: Of 494 donors included, 432 (87·4%) were enrolled from fixed blood collection sites and 62 (12·6%) were contacted by phone. A total of 464 (93·9%) participants believed that the COVID-19 is a lung disease, but some declared it to be imaginary (7·7%), a foreign disease (8·3%) or a blood-borne disease (3·2%). The participants reported that the distribution of face masks and hydroalcoholic solutions (92·5%), social distancing and hygiene (6·3%) are the most important measures that need to be in place for safe donation. The number of blood donations dropped by 21·5% between 2019 and 2020. CONCLUSION: Most of the donors know COVID-19, its transmission routes and manifestations. In the absence of barrier measures, they perceive blood donation as a threat to their health. Distribution of masks and hydroalcoholic solution might motivate more donors and improve the blood supply.

Prevalence and factors associated with overweight and obesity in selected health areas in a rural health district in Cameroon: a cross-sectional analysis.

BACKGROUND: Overweight and obesity are major public health problems worldwide, with projections suggesting a proportional increase in the number of affected individuals in developing countries by the year 2030. Evidence-based preventive strategies are needed to reduce the burden of overweight and obesity in developing countries. We assessed the prevalence of, and factors associated with overweight and obesity in selected health areas in West Cameroon. METHODS: Data were collected from a community-based cross-sectional study, involving the consecutive recruitment of participants aged 18 years or older. Overweight and obesity were defined according to the WHO classification. The statistical software R (version 3.5.1, The R Foundation for statistical computing, Vienna, Austria) was used for statistical analysis. Multivariable logistic regression analysis was used to assess independent factors associated with overweight and obesity, and obesity. RESULTS: Records of 485 participants were included for analysis. The age and sex-standardized prevalence of overweight, obesity, and overweight and obesity were 31.1% (95% CI, 27.0-35.2), 18.9% (95% CI, 14.9-22.9), and 50.1% (95% CI, 45.7-54.6), respectively. In multivariable analysis, being female (adjusted OR [aOR] = 2.79, 95% CI = 1.69-4.63), married (aOR = 3.90, 95% CI = 2.23-6.95), and having secondary or tertiary education (aOR = 3.27, 95% CI = 1.77-6.17) were associated with higher odds of overweight and obesity, while current smokers had lower odds of overweight and obesity (aOR = 0.37, 95% CI = 0.16-0.82) when compared to their respective counterpart. Compared to their respective reference categories, being female being (aOR = 3.74, 95% CI = 2.01-7.30), married (aOR = 2.58, 95% CI = 1.37-5.05) and having secondary or tertiary education (aOR = 2.03, 95% CI = 1.00-4.23) were associated with higher odds of obesity after adjustments for confounding. CONCLUSION: We observed a high prevalence of overweight and obesity in this study. The odds of overweight and obesity was higher in females, married participants, and those with higher levels of education. Community-based interventions to control overweight and obesity should consider targeting these groups.

The use of the GeeniusTM HIV-1/2 Rapid confirmatory test for the enrolment of patients and blood donors in the WHO Universal Test and Treat Strategy in Cameroon, Africa.

BACKGROUND AND OBJECTIVE: In the WHO Universal test and treat strategy, false-positive HIV blood donors and patients may be unnecessarily put under antiretroviral treatment and false-negative subjects may be lost to follow-up. This study assessed the false positivity rate of the Cameroonian national HIV screening testing algorithm and the benefit of a confirmation test in the enrolment of patients and donors in the HIV care programme. METHODS: We included initial HIV reactive blood donors and patients in a cross-sectional study conducted in two Cameroonian hospitals. Samples were retested according to the Cameroon national algorithm for HIV diagnosis. A positive or discordant sample was retested with the Geenius Bio-Rad HIV 1&2 (Bio-Rad, Marnes-la-Coquette, France) for confirmation. The Geenius HIV-1-positive results with 'poor' profiles were retested for RNA as well as the Geenius indeterminate results. RESULTS: Of the 356 participants, 190/225 (84·4%) patients and 76/131 (58%) blood donors were declared positive with the national algorithm; 257 participants (96·6%) were confirmed HIV-1-positive. The study revealed that about 34/1000 blood donors and patients are false-positive and unnecessarily put on treatment; 89/1000 blood donors and patients declared discordant could have been included immediately in the HIV care programme if confirmatory testing was performed. The second test of the algorithm had a false-negative rate of 3%. Eleven samples (3·1%) were Geenius poor positive and NAT negative. CONCLUSION: The universal test and treat strategy may identify and refer more individuals to HIV care if a third rapid confirmatory test is performed for discordant cases.

Attention/Working Memory, Learning and Memory in Adult Cameroonians: Normative Data, Effects of HIV Infection and Viral Genotype.

OBJECTIVE: There is lack of Cameroonian adult neuropsychological (NP) norms, limited knowledge concerning HIV-associated neurocognitive disorders in Sub-Saharan Africa, and evidence of differential inflammation and disease progression based on viral subtypes. In this study, we developed demographically corrected norms and assessed HIV and viral genotypes effects on attention/working memory (WM), learning, and memory. METHOD: We administered two tests of attention/WM [Paced Auditory Serial Addition Test (PASAT)-50, Wechsler Memory Scale (WMS)-III Spatial Span] and two tests of learning and memory [Brief Visuospatial Memory Test-Revised (BVMT-R), Hopkins Verbal Learning Test-Revised (HVLT-R)] to 347 HIV+ and 395 seronegative adult Cameroonians. We assessed the effects of viral factors on neurocognitive performance. RESULTS: Compared to controls, people living with HIV (PLWH) had significantly lower T-scores on PASAT-50 and attention/WM summary scores, on HVLT-R total learning and learning summary scores, on HVLT-R delayed recall, BVMT-R delayed recall and memory summary scores. More PLWH had impairment in attention/WM, learning, and memory. Antiretroviral therapy (ART) and current immune status had no effect on T-scores. Compared to untreated cases with detectable viremia, untreated cases with undetectable viremia had significantly lower (worse) T-scores on BVMT-R total learning, BVMT-R delayed recall, and memory composite scores. Compared to PLWH infected with other subtypes (41.83%), those infected with HIV-1 CRF02_AG (58.17%) had higher (better) attention/WM T-scores. CONCLUSIONS: PLWH in Cameroon have impaired attention/WM, learning, and memory and those infected with CRF02_AG viruses showed reduced deficits in attention/WM. The first adult normative standards for assessing attention/WM, learning, and memory described, with equations for computing demographically adjusted T-scores, will facilitate future studies of diseases affecting cognitive function in Cameroonians.

Malaria in patients with sickle cell anaemia: burden, risk factors and outcome at the Laquintinie hospital, Cameroon.

BACKGROUND: It is believed that the current prevalence of malaria in endemic areas reflects selection for the carrier form of sickle cell trait through a survival advantage. Malaria has been incriminated as a great cause of mortality in people with sickle cell disease (SCD). However, people with SCD, a high-risk group, do not benefit from free or subsisized malaria prevention and treatment in Cameroon unlike other vulnerable groups which may be due to insufficient evidence to guide policy makers. This study aimed at describing clinical and socio-demographic characteristics of patients with malaria, determining the prevalence of malaria in hospitalized children and in those with SCD and without, compare frequency of presentation of malaria related complications (using clinical and laboratory elements that define severe malaria) between children admitted for malaria with SCD and those without and finally, determing the risk factors for death in children admitted for malaria. METHODS: This was a retrospective analysis of admission records of children age 1 to 18 years with a confirmed malaria diagnosis admitted at the Laquintinie Hospital during January 2015 through December 2018. Clinical features, laboratory characteristics and outcome of malarial infections, stratified by SCD status were studied. Patients with HIV infection, malnutrition, renal failure and discharged against medical advice were excluded from the study. Data were analysed using Epi-info 7 software and analysis done. Chi square test, Odds ratios, CI and student's t test were used to determine association between variables. Statistical significance was set at p-value ≤0.05. RESULTS: The prevalence of malaria was lower among children with SCD than it was among children without SCD (23.5% vs 44.9%). Similarly, among those with a positive microscopy, the mean parasite density was significantly lower among children with SCD than it was among children without SCD (22,875.6 vs 57,053.6 parasites/ µl with t-value - 3.2, p-value 0.002). The mean hemoglobin concentration was lower in SCD as compared to non SCD (5.7 g/l vs 7.4 g/l, t-value - 12.5, p-value < 0.001). Overall mortality in SCD was 3.4% and malaria was reponsible for 20.4% of these deaths as compared to the 35.4% in non SCD patients. Convulsion and impaired consciousness were significantly lower in SCD group (OR:0.1, CI: 0.1-0.3, p value < 0.01 and OR:0.1, CI:0.1-0.2, p-value < 0.001 respectively). Death was significantly higher in SCD patients with malaria as compared to SCD patients admitted for other pathologies (3.2% vs 1.5%., OR:2.2, CI:1-5, p-value 0.050). CONCLUSION: The SCD population has a lower mortality related to malaria compared to the non-SCD population. Meanwhile, within the SCD population, those admitted with malaria are twice more likely to die than those admitted for other pathologies. Jaundice, hepatomegaly and splenomegaly were common in SCD with malaria, however no risk factors for malaria severity or malaria related death was identified.

Programme quality indicators of HIV drug resistance among adolescents in urban versus rural settings of the centre region of Cameroon.

BACKGROUND: The high rate of mortality among HIV-vertically infected adolescents might be favoured by HIV drug resistance (HIVDR) emergence, which calls for timeous actions in this underserved population. We thus sought to evaluate program quality indicators (PQIs) of HIVDR among HIV-vertically infected adolescents on antiretroviral therapy (ART). METHODS: A study was conducted in the Centre region of Cameroon among adolescents (10-19 years) receiving ART in two urban (The Mother-Child Centre of the Chantal BIYA Foundation, the National Social Welfare Hospital) and three rural (Mfou District Hospital, Mbalmayo District Hospital and Nkomo Medical Center) health facilities. Following an exhaustive sampling from ART registers, patient medical files and pharmacy records, data was abstracted for seven PQIs: on-time drug pick-up; retention in care; pharmacy stock outs; dispensing practices; viral load coverage; viral suppression and adequate switch to second-line. Performance in PQIs was interpreted following the WHO-recommended thresholds (desirable, fair and/or poor); with p < 0.05 considered significant. RESULTS: Among 967 adolescents (888 urban versus 79 rural) registered in the study sites, validated data was available for 633 (554 in urban and 79 in rural). Performance in the urban vs. rural settings was respectively: on-time drug pick-up was significantly poorer in rural (79% vs. 46%, p = 0.00000006); retention in care was fair in urban (80% vs. 72%, p = 0.17); pharmacy stock outs was significantly higher in urban settings (92% vs. 50%, p = 0.004); dispensing practices was desirable (100% vs. 100%, p = 1.000); viral load coverage was desirable only in urban sites (84% vs. 37%, p < 0.0001); viral suppression was poor (33% vs. 53%, p = 0.08); adequate switch to second-line varied (38.1% vs. 100%, p = 0.384). CONCLUSION: Among adolescents on ART in Cameroon, dispensing practices are appropriate, while adherence to ART program and viral load coverage are better in urban settings. However, in both urban and rural settings, pharmacy stock outs, poor viral suppression and inadequate switch to second-line among adolescents require corrective public-health actions to limit HIVDR and to improve transition towards adult care in countries sharing similar programmatic features.

Variability in HIV-1 Integrase Gene and 3'-Polypurine Tract Sequences in Cameroon Clinical Isolates, and Implications for Integrase Inhibitors Efficacy.

: Integrase strand-transfer inhibitors (INSTIs) are now included in preferred first-line antiretroviral therapy (ART) for HIV-infected adults. Studies of Western clade-B HIV-1 show increased resistance to INSTIs following mutations in integrase and nef 3'polypurine tract (3'-PPT). With anticipated shifts in Africa (where 25.6-million HIV-infected people resides) to INSTIs-based ART, it is critical to monitor patients in African countries for resistance-associated mutations (RAMs) affecting INSTIs efficacy. We analyzed HIV-1 integrase and 3'-PPT sequences in 345 clinical samples from INSTIs-naïve HIV-infected Cameroonians for polymorphisms and RAMs that affect INSTIs. Phylogeny showed high genetic diversity, with the predominance of HIV-1 CRF02_AG. Major INSTIs RAMs T66A and N155K were found in two (0.6%) samples. Integrase polymorphic and accessory RAMs found included T97A, E157Q, A128T, M50I, S119R, L74M, L74I, S230N, and E138D (0.3%-23.5% of samples). Ten (3.2%) samples had both I72V+L74M, L74M+T97A, or I72V+T97A mutations; thirty-one (9.8%) had 3'-PPT mutations. The low frequency of major INSTIs RAMs shows that INSTIs-based ART can be successfully used in Cameroon. Several samples had 1 INSTIs accessory RAMs known to reduce INSTIs efficacy; thus, INSTIs-based ART would require genetic surveillance. The 3'-PPT mutations could also affect INSTIs. For patients failing INSTIs-based ART with no INSTIs RAMs, monitoring 3'-PPT sequences could reveal treatment failure etiology.

Hepatitis C virus surveillance and identification of human pegivirus 2 in a large Cameroonian cohort.

The prevalence of chronic hepatitis C virus (HCV) and the presence of human pegivirus 2 (HPgV-2) have not been examined in Cameroon, although HCV has been associated with HPgV-2 infections previously. Herein we aimed to characterize the burden and genetic diversity of HCV and the presence of HPgV-2 in Cameroon. Retrospective plasma specimens collected from N = 12 369 consenting subjects in South Cameroon from 2013 to 2016 were included in the study. The majority (97.1%) of participants were patients seeking health care. All specimens were screened for HCV using the Abbott RealTime HCV viral load assay and positive specimens with remaining volume were also screened for HPgV-2 antibodies on the Abbott ARCHITECT instrument, followed by molecular characterization. Overall, HCV RNA was detected in 305 (2.47%; 95% CI: 2.21%-2.75%) specimens. Notably, the prevalence of HCV RNA was 9.09% amongst participants over age 40 and 3.81% amongst males. Phylogenetic classification of N = 103 HCV sequences identified genotypes 1 (19.4%), 2 (15.5%) and 4 (65.1%) within the study cohort. Amongst HCV RNA-positive specimens, N = 28 (10.6%; 95% CI: 7.44%-14.90%) specimens also had detectable HPgV-2 antibodies. Of these, N = 2 viremic HPgV-2 infections were confirmed by sequencing and shared 93-94 median % identity with strains found on other continents. This is the first study to determine the prevalence of chronic HCV in Cameroon, and the discovery of HPgV-2 in this study cohort expands the geography of HPgV-2 to the African continent, indicating a widespread distribution exists.

Risk factors for human immunodeficiency virus among blood donors in Cameroon: evidence for the design of an Africa-specific donor history questionnaire.

BACKGROUND: In sub-Saharan Africa improving the deferral of at-risk blood donors would be a cost-effective approach to reducing transfusion-transmitted human immunodeficiency virus (HIV) infections. We performed a pilot case-control study to identify the risk factors for HIV infection and to develop an adapted donor history questionnaire (DHQ) for sub-Saharan Africa. STUDY DESIGN AND METHODS: We recruited 137 HIV-positive donors (cases) and 256 HIV-negative donors (controls) and gathered risk factor data using audio computer-assisted self-interview. Variables with univariate associations were entered into a logistic regression model to assess independent associations. A scoring scheme to distinguish between HIV-positive and HIV-negative donors was developed using receiver operating characteristics curves. RESULTS: We identified 16 risk factors including sex with sex worker, past history or treatment for sexually transmitted infections, and having a partner who used injected or noninjected illegal drugs. Two novel risks were related to local behavior: polygamy (odds ratio [OR], 22.7; 95% confidence interval [CI], 5.9-86.7) and medical or grooming treatment on the street (OR, 1.8; 95% CI, 1.0-3.0). Using the 16 selected items the mean scores (>100) were 82.6 ± 6.7 (range, 53.2-95.1) and 85.1 ± 5.2 for HIV-negative donors versus 77.9 ± 6.8 for HIV-positive ones (p = 0.000). Donors who scored between 80 and 90 were more likely to be HIV negative than those who scored less (OR, 31.4; 95% CI, 3.1-313.9). CONCLUSION: We identified both typical and novel HIV risk factors among Cameroonian blood donors. An adapted DHQ and score that discriminate HIV-negative donors may be an inexpensive means of reducing transfusion-transmitted HIV through predonation screening.

Psychosocial burden of sickle cell disease on parents with an affected child in Cameroon.

The chronicity of Sickle Cell Disease (SCD) could impair the quality of life of caregivers. We performed a quantitative study to assess various indices of psychosocial burden on Cameroonian parents (N = 130) with at least one living SCD-affected child. Demographic and medical information were obtained from the participants and the review of the patients' medical records. The survey instrument included a 38-item stress factors scale using Likert-type statements, evaluating general perceptions of stress and five main specific stressors: disease factors (clinical severity), hospital factors, financial factors, family factors (life/dynamic) and SCD-child factors (perceived quality of life). The items pertaining to burden involved four response options with increasing severity: 0, 1, 2 or 3. Descriptive statistics and non-parametric tests were used for analysis. Participants were typically aged 38 years, urban dwellers (89%), female (80%), married (60.2%), employed (61.7%) and had secondary/tertiary education (82%). Median age of SCD-affected children was 9 years. The median age at diagnosis of SCD was 6 months; 47.8% had more than 3 painful crises per year. The majority of participants (88.3%) experienced moderate to severe difficulty coping with SCD. On a 0-3 scale, median score of SCD clinical severity was the major factor to undermine the coping ability of parents (2.2); vaso-occlusive painful events (>3 per year) was the disease-related stressor that most impacted their coping ability. The family life dynamic was the least stressful (0.7). Unemployment affected all the stressors' categories. Stressors scores also increased with female, single, low education level, age of SCD-affected children or more than 3 children in the family. In Cameroon, there is an urgent need to implement practices that ensure affordable access to health-care and activities that would reduce SCD morbidity.

Psychosocial stressors of sickle cell disease on adult patients in Cameroon.

Sickle Cell Disease (SCD) is a debilitating illness that affects quality of life. Studies of the psychosocial burden of SCD on patients have been rarely reported in Africa. We used a quantitative method, with face-to-face administered questionnaires, to study indices of psychosocial stressors on adult SCD patients in Cameroon. The questionnaire included a 36-item stress factors scale evaluating general perceptions of stress and five main stressors' domain: disease factors, hospital factors, financial factors, family factors and quality of personal-life factors. Items pertaining to psychosocial stressors involved four response options with increasing severity: 0, 1, 2 or 3. Non-parametric tests were used for analysis. The majority of the 83 participants were urban dwellers, female, 20-30 years old, single, unemployed, with at least a secondary or tertiary education. Median age at diagnosis was 100 months; 47.8% had >3 painful vaso-oclusive crises annually. Only 4.8% had been treated with hydroxyurea. The majority reported moderate to severe difficulty coping with SCD. The "degree of clinical severity" category displayed the highest median score (2.0), while familial stressors showed the lowest (0.8). Being female, married, with low education level, an additional affected sibling and low direct income were significantly associated with specific stressors' categories. In Cameroon, there is an urgent need to implement policies that ensure affordable access to health-care and practices to reduce SCD morbidity and improve patients' quality of life.

Evaluation of a fully automated high-throughput serology assay for detection of Hepatitis D virus antibodies.

BACKGROUND: HDV antibody testing is recommended for universal screening and as the first line in an HDV double reflex testing strategy for effectively identifying patients with active infection for therapeutic treatments. OBJECTIVE: The aim of this study is to evaluate the performance of a newly developed ARCHITECT HDV Total Ig (ARCHITECT HDV Ig) prototype assay. STUDY DESIGN: Performance characteristics were determined for the ARCHITECT HDV Ig and a reference test, LIAISON XL Anti-HDV using a well-characterized specimen panel, comprising HDV RNA positive (n = 62) and negative (n = 70) samples, and healthy US blood donors. RESULTS: Healthy US blood donors (n=200) showed 99.5% (199/200, 95%CI=97.65-99.98) specificity with ARCHITECT HDV Ig and 98.5 % (197/200, 95 %CI = 96.10-99.64) with LIAISON Anti-HDV. Among known HDV RNA positive samples, ARCHITECT HDV Ig detected 59/62 demonstrating 95.2 % sensitivity while LIAISON Anti-HDV sensitivity was 90.3 % (56/62). Among 101 HBV positive samples, 70 were reactive in the ARCHITECT test, 59 of which tested positive for HDV RNA for a positive predictive value (PPV) for the presence of HDV RNA was 84.3 %. For LIAISON Anti-HDV, 79 specimens were reactive and 56 contained HDV RNA: PPV for HDV RNA was 70.9 %. Among 70 HDV RNA negative samples, 39 were HBV positive. ARCHITECT HDV Ig negative predictive value (NPV) was 71.8 % and LIAISON Anti-HDV NPV was 41 % for the HBV positive group, respectively. CONCLUSION: When compared to the LIASON Anti-HDV test, the ARCHITECT HDV Ig assay demonstrated enhanced sensitivity and specificity and better NPV and PPV values for HDV RNA status. The ARCHITECT HDV Ig assay represents a promising tool for universal screening of all HBsAg-positive persons.

Optimizing Haemophilia Care in Resource-Limited Countries: Current Challenges and Future Prospects.

About 75% of persons with hemophilia live in the developing world and do not have access to routine care due to many barriers. There are a lot of challenges associated with hemophilia care in resource-limited settings, ranging from financial to organisational and government commitments. This review discusses some of these challenges and future prospects, while highlighting the important role of the World Federation of Hemophilia in hemophilia patient care. A participative approach involving all stakeholders is key to optimizing care in resource-limited settings.

Measuring hepatitis B pgRNA stability using an updated automated HBV pgRNA assay with increased sensitivity.

BACKGROUND: HBV pregenomic RNA (pgRNA) is a circulating biomarker for covalently closed circular DNA activity in HBV-infected individuals and has been studied for treatment efficacy, disease staging, and off-therapy outcomes; however, data on the stability are scarce. Increasing HBV pgRNA assay sensitivity may improve its predictive value and provide additional insights at low viral levels. METHODS: Modifications to a fully automated first (v1) generation HBV pgRNA assay improved sensitivity up to 15-fold over the previous assay. Flexible sample input volumes yielded lower limits of quantitation of 10 and 22 copies/mL for 0.6 and 0.2 mL assays, respectively. Results are standardized to secondary standards that are traceable to the WHO HBV DNA standard, and internal and external controls are included. RESULTS: Comparison between v1 and modified v2 assays showed increased sensitivity from 152 copies/mL with v1 to 10 (0.6 mL) and 22 (0.2 mL) copies/mL with v2, respectively. Quantitated v2 results were indistinguishable from v1, indicating that comparisons can be made to previous studies. Single timepoint treatment-naive blood donors or longitudinal draws from patients with chronic hepatitis B on AB-729, an investigational siRNA therapy, showed improved detection and quantifiable pgRNA with v2 compared with v1. Stability testing demonstrated excellent HBV pgRNA plasma stability after 3 freeze-thaw cycles, for at least 7 days at 25-37 °C and at least 30 days at 4°C, with ≤0.25 Log U/mL decrease. CONCLUSION: HBV pgRNA v2 assays with increased sensitivity and flexible input volumes demonstrated increased detection and quantitation of low viral titer samples. Highly sensitive HBV pgRNA assays may be useful in refining predictive treatment outcomes based on this marker. HBV pgRNA was stable under multiple conditions, which increases the reliability of this marker.

Metagenomic Detection of Divergent Insect- and Bat-Associated Viruses in Plasma from Two African Individuals Enrolled in Blood-Borne Surveillance.

Metagenomic next-generation sequencing (mNGS) has enabled the high-throughput multiplexed identification of sequences from microbes of potential medical relevance. This approach has become indispensable for viral pathogen discovery and broad-based surveillance of emerging or re-emerging pathogens. From 2015 to 2019, plasma was collected from 9586 individuals in Cameroon and the Democratic Republic of the Congo enrolled in a combined hepatitis virus and retrovirus surveillance program. A subset (n = 726) of the patient specimens was analyzed by mNGS to identify viral co-infections. While co-infections from known blood-borne viruses were detected, divergent sequences from nine poorly characterized or previously uncharacterized viruses were also identified in two individuals. These were assigned to the following groups by genomic and phylogenetic analyses: densovirus, nodavirus, jingmenvirus, bastrovirus, dicistrovirus, picornavirus, and cyclovirus. Although of unclear pathogenicity, these viruses were found circulating at high enough concentrations in plasma for genomes to be assembled and were most closely related to those previously associated with bird or bat excrement. Phylogenetic analyses and in silico host predictions suggested that these are invertebrate viruses likely transmitted through feces containing consumed insects or through contaminated shellfish. This study highlights the power of metagenomics and in silico host prediction in characterizing novel viral infections in susceptible individuals, including those who are immunocompromised from hepatitis viruses and retroviruses, or potentially exposed to zoonotic viruses from animal reservoir species.

Characterization of HBV surface antigen isoforms in the natural history and treatment of HBV infection.

BACKGROUND: The loss of HBV HBsAg or functional cure is a desirable goal of hepatitis B management. The relative abundances of HBsAg isoforms may offer additional diagnostic and predicting values. To evaluate the clinical utility of HBsAg isoforms, we developed novel prototype assays on the ARCHITECT automated serology platform that specifically detects total-HBsAg (T-HBsAg), large (L-HBsAg), and middle (M-HBsAg) products of the S gene to determine the isoform composition of human specimens from acute and chronic HBV infection and during long-term nucleos(t)ide analog therapy. RESULTS: In the early phase of acute HBV infection, L-HBsAg and M-HBsAg emerged within days and were in parallel to T-HBsAg during the entire course of infection. M-HBsAg levels were consistently higher than L-HBsAg levels. Patients with HBeAg(+) chronic hepatitis B had higher T-HBsAg, M-HBsAg, and L-HBsAg levels compared with HBeAg(-) patients. Correlations of M-HBsAg and L-HBsAg to T-HBsAg were similar in both. In contrast, there was no strong correlation between L-HBsAg or M-HBsAg with HBV DNA levels. During long-term nucleos(t)ide analog treatment, changes in HBsAg isoform abundance were proportional to T-HBsAg regardless of treatment responses for both HBeAg(+) and HBeAg(-) chronic hepatitis B. A larger sample size may be necessary to detect a significant difference. CONCLUSION: HBsAg isoform compositions parallel T-HBsAg levels in both acute and chronic hepatitis B infection. L-HBsAg and M-HBsAg individual biomarkers do not appear to provide an additional diagnostic benefit for staging chronic disease or monitoring response to treatment with current therapies.