Virology unit

Hepatitis Viruses and Other Oncogenic Viruses

Liver Cancer in Equatorial Africa: Characterization of New Serum Biomarkers of Primary Liver Cancer in Patients in Equatorial Africa (Prof. Njouom)

Objective: This study aims to identify its infectious and non-infectious etiological factors; to evaluate the influence of different viral genotypes and variants on its development; and to describe the somatic genetics of liver tumors.

Progress Status

To date, 88 cases of hepatocellular carcinoma (HCC), 85 control group 1, and 140 control group 2 (100 for HBV and 40 for HCV) have been included in this study. Serological analyses of HBV and HCV among cases and controls revealed that both viruses are strongly associated with the development of HCC in Cameroon, with respective prevalences of (64% and 26%) among HCC subjects versus (10% and 4.3%) among controls without HCC. Phylogenetic analyses of HBV revealed the circulation of two genotypes,

A and E, in our study population, with 35/42 genotype A (83.3%) and 7/42 genotype E (16.7%) among cases, compared to 13/44 genotype A (29.5%) and 31/44 genotype E (70.5%) among controls. Phylogenetic analyses of HCV revealed the circulation of three genotypes—1, 2, and 4—in our study population, with 7/17 genotype 4 (41.2%), 5/17 genotype 2 (29.4%), and 5/17 genotype 1 (29.4%) among cases, versus 2/8 genotype 1 (25%), 4/8 genotype 2 (50%), and 2/8 genotype 4 (25%) among controls. Evaluation of HDV serology among all patients positive for HBsAg showed that among 54 HCC cases tested, 21 (38.9%) were positive, while among healthy controls without cancer, only 1/6 (16.7%) tested positive for anti-HDV antibodies.

Mutations in the Pre-S and C genes have already been identified. The mutations C1766T, T1768A, and G1862T/A in the HBV C gene were found only in cases, whereas mutations C2875A, G2951C, and C3116T in the HBV pre-S fragment were significantly more frequent among cases compared to HBV-positive controls. These mutations may therefore be strongly involved in the development of HCC among HBV-infected patients.

Epidemiology of Viral Hepatitis B, C, and Delta in Cameroon: Analysis of Samples from the 2011 Demographic and Health Survey (DHS Hepatitis). ANRS 12289 Project (Prof. Njouom)

Objectives: Given the availability of data and the biobank from the 2011 Cameroon National Demographic and Health Survey (DHS), this project was initiated to study the seroprevalence, distribution, and factors associated with HBV, HCV, and HDV infections in a representative sample of the general population, and to determine the contribution of historical iatrogenic HCV transmission to the current epidemic in Cameroon.

Activities Conducted and Conclusions

Analyses yielded the following national prevalence rates: Hepatitis B Virus: 11.9% (95% CI: 11.0%–12.8%); Hepatitis C Virus: 1.03% (95% CI: 0.8%–1.6%); Hepatitis D Virus: 10.5% (95% CI: 8.7%–12.7%).

These results show that Cameroon may be considered a low-endemic area for hepatitis C and a high-endemic area for hepatitis B and D. Prevalence varies widely across the country, indicating distinct epidemiological patterns in certain administrative regions.

HBV prevalence differs significantly by age group: between 11–13% among individuals under 45 years, and between 8–9% among those over 45 (p<0.05). Rural populations are more affected than urban ones (13.3% vs. 10.7%, p<0.05). Prevalence also differs significantly by administrative region, with the Far North and North regions most affected (17.7% and 17.5%, respectively).

Among HBV-infected individuals, HDV prevalence also varies significantly by age, with those over 45 years being the most affected (16.8%). HDV prevalence also differs by region, with the East and South regions showing the highest rates (53.7% and 48.5%, respectively).

HCV prevalence increases significantly with age—from about 1% in those under 45, to over 3% among those aged 45–55, and about 7% among those over 55. A recent study in Ebolowa among individuals over 60 found a prevalence of about 50% in this population group (Pépin et al., 2010). Rural populations are more affected than urban ones (1.2% vs. 0.9%, p<0.05). Prevalence also varies by region, with the Centre region being the most affected (2.4%).

In summary, at-risk populations vary depending on the type of hepatitis virus: Hepatitis B among young people under 45 years (prevalence above 10%), and Hepatitis C among older adults over 55 years (prevalence around 7%).

Strategies to combat each of these viral hepatitis infections should consider these at-risk populations: Hepatitis B among the young, and Hepatitis C among the elderly. Observational study on the effectiveness of hepatitis B (HB) vaccination initiated at birth and follow-up of the expanded vaccination program including the EPI-HB component among children born to HBsAg-positive mothers in the Tokombéré district, Cameroon. ANRS 12303 (Prof. Richard Njouom)

Objectives:

To evaluate the effectiveness of “targeted prevention of mother-to-child transmission (PMTCT) of HBV,” initiated since 2009 in Tokombéré with GEMHEP support, involving systematic screening during pregnancy for HIV and HBsAg, followed by HB vaccination at birth for infants born to HBsAg-positive mothers (without anti-HBs immunoglobulin), and subsequent administration of the three EPI-HB doses at 6, 10, and 14 weeks.

The primary objective is to evaluate vaccination effectiveness in the target population by estimating the mother-to-child transmission rate among infants vaccinated at birth from HBsAg-positive mothers (HB birth vaccination failures).

The secondary objective is to study the risk factors associated with these failures (e.g., vaccination delay).

Project Progress Summary

Recruitment of mother-child pairs is ongoing in Tokombéré. Blood samples are collected and will be sent to CPC and the University Hospital of Angers, France, for HBV serological and molecular analyses. However, security concerns in the Far North of Cameroon have slowed recruitment.

Evaluation of the Performance of Three Rapid Diagnostic Tests for the Detection of Anti-Hepatitis C Virus Antibodies in Cameroon.

Acronym: (ANRS HEPATITE 12336) (Prof. Richard Njouom)

Research Objective

– To evaluate the performance of three rapid diagnostic tests (Multisure HCV Antibody Assay, First Response® HCV Card Test, and Toyo® Anti-HCV Test) for detecting anti-HCV antibodies in Cameroon.

– To determine sensitivity, specificity, predictive values (positive and negative), likelihood ratios (positive and negative), and the area under the ROC curve for these three RDTs using plasma samples.

– To determine the same performance indicators for the Multisure test using whole blood dried on filter paper (DBS).

Progress Status:

A retrospective cross-sectional study was conducted at the Centre Pasteur du Cameroun (CPC) over six months (July–December 2016). Using the ABBOTT Diagnostic Architect anti-HCV immunoassay as reference, a panel of 200 plasma samples (100 positive and 100 negative) was assembled from the CPC plasmathèque. The three RDTs (Multisure, First Response®, and Toyo®) were evaluated on plasma. DBS were reconstituted from 200 whole blood samples (100 negative and 100 positive) from patients who came to CPC for HCV serology. Only the Multisure test was evaluated on DBS. Test performance was expressed as sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (LR+), negative likelihood ratio (LR–), and area under the ROC curve (AUC).

Evaluation on plasma yielded: Se (99%, 96%, 96%), Sp (83%, 90%, 78%), PPV (85.3%, 96%, 89.7%), NPV (98.8%, 90%, 90.7%), LR+ (5.82, 9.6, 4.3), LR– (0.01, 0.04, 0.05), and AUC (0.96, 0.93, 0.88) for Multisure, First Response®, and Toyo®, respectively. On DBS, Multisure yielded: Se (71%), Sp (100%), PPV (100%), NPV (78.7%), LR+ (∞), LR– (0.27), and AUC (0.68).

Diagnosis and Monitoring of Viral Hepatitis

Epidemiology of Viral Hepatitis B, C, and Delta in Cameroon: Analysis of Samples from the 2011 Demographic and Health Survey (DHS) ANRS 12289-Hepatitis C (Prof. Njouom)

Objectives: Using data and biobank resources from the 2011 Cameroon DHS, we initiated this project to study seroprevalence, distribution, and factors associated with HBV, HCV, and HDV infections in a representative population sample and to determine the contribution of historical iatrogenic HCV transmission to the current epidemic.

Progress: All biological analyses (HIV, HCV, HBV, and HDV serology) were completed on over 14,000 samples collected during DHS-4. Statistical analysis is ongoing.

Diagnosis and Management of Chronic Viral Hepatitis

The high prevalence of chronic HBV and HCV infection in Northern Cameroon makes patient management a daily challenge for clinicians. While diagnostic tests for viral carriage are accessible, access to tests confirming viral replication, genotype, and disease stage is limited. These essential tests for appropriate therapy are expensive and must be prescribed judiciously. Dr. Richard Njouom, Head of the Virology Service at the Centre Pasteur du Cameroun in Yaoundé, organized a meeting in Garoua, Cameroon, on November 3, 2015, to address practical issues such as: How to diagnose chronic viral hepatitis? What assessments should follow diagnosis? How to initiate antiviral treatment? What biological follow-up should be offered to patients?

Prevalence and Genetic Diversity of Hepatitis Viruses

Prevalence and Genetic Diversity of Hepatitis E Virus in Porcine and Human Populations in Cameroon: Evaluation of interspecies transmission. This project, coordinated by Prof. Richard Njouom, is funded by the University of California, Los Angeles.

Epidemiology of Viral Hepatitis B, C, and Delta in Cameroon: Analysis of samples from the 2011 Demographic and Health Survey. ANRS Project.

This project is a collaboration with the Institut Pasteur (Prof. Arnaud Fontanet) and the Universities of Sherbrooke, Canada (Dr. J. Pépin) and Paris Diderot (G. Lachenal).

The demonstration of effective therapeutic protocols and the introduction of affordable generic or biosimilar drugs are expected to revolutionize hepatitis B and C management in developing countries. It is now urgent for the most affected countries to obtain reliable estimates of the number of infected individuals who will require treatment in the coming years.

Cameroon is one of the Central African countries most affected by chronic viral hepatitis, especially hepatitis C. The data and biobank from the 2011 Cameroon DHS provide a unique opportunity to study the seroprevalence, distribution, and factors associated with HBV, HCV, and HDV infections in a representative general population sample. The project also aims to identify factors associated with HCV infection and assess the contribution of historical iatrogenic transmission to the current epidemic.

HBV, HCV, and HDV serology as well as HBV DNA quantification will be performed on 14,700 dried blood spot (DBS) samples collected during the 2011 DHS. Historical iatrogenic transmission of HCV will be analyzed through geo-referenced DHS data and spatial correlation analyses between HCV infection and historical maps of medical interventions.

Molecular Diagnosis of Human Papillomavirus (HPV)

HPVs (Human Papillomaviruses) are viruses that can infect the skin and mucous membranes. There are more than 150 types of HPV, about 40 of which can infect male and female genital organs. Around 20 types can cause moderate to severe cellular abnormalities, precancerous lesions, and cancers. In collaboration with the Department of Anatomopathology, the Virology Service has implemented molecular diagnosis of HPV to confirm suspected cases detected by cervical-vaginal smears during cervical cancer screening. In 2012, 89 molecular diagnostic tests (by viral DNA detection on cervical smears) were performed.