Search

The co-circulation of West Nile virus (WNV), Usutu virus (USUV), and tick-borne encephalitis virus (TBEV) in Europe is a public health concern, and the lack of active surveillance programs and high-quality specific serology tests make it difficult to estimate the true disease burden. The potential for introducing other pathogens, such as the related Japanese encephalitis virus (JEV), could further complicate this picture. The standard method for diagnosing flavivirus infections involves RT-(q)PCR to detect the viral RNA during the acute phase of the disease, followed by antibody detection in the convalescence phase. Both methods suffer from several limitations that make flavivirus diagnosis challenging, especially in areas of co-circulation. Non-structural protein 1 (NS1) represents a promising marker for discriminating between co-circulating flaviviruses. However, NS1 antigen-capture tests are lacking on the commercial market for WNV, USUV, TBEV, and JEV. In recent decades, the presence of flaviviruses of concern for human health in Europe has drastically increased,exacerbated by the effects of climate change – which has allowed the vectors of these viruses to expand into new territories. Co-circulation of West Nile virus (WNV), Usutu virus (USUV), and tick-borne encephalitis virus (TBEV) represents a threat to the European continent, and this is further complicated by the difficulty of obtaining an early and discriminating diagnosis of infection. Moreover, the possibility of introducing non-endemic pathogens, such as Japanese encephalitis virus (JEV), further complicates accurate diagnosis. Current flavivirus diagnosis is based mainly on RT-PCR and detection of virus-specific antibodies. Yet, both techniques suffer from limitations, and the development of new assays that can provide an early, rapid, low-cost, and discriminating diagnosis of viral infection is warranted. In the pursuit of ideal diagnostic assays, flavivirus non-structural protein 1 (NS1) serves as an excellent target for developing diagnostic assays based on both the antigen itself and the antibodies produced against it. This review describes the potential of such NS1-based diagnostic methods, focusing on the application of flaviviruses that co-circulate in Europe. [ABSTRACT FROM AUTHOR]

Chikungunya virus (CHIKV) is an arthropod-borne virus that has re-emerged recently and has spread to previously unaffected regions, resulting in millions of infections worldwide. The genus Alphavirus , in the family Togaviridae, contains several members with a similar potential for epidemic emergence. In order for CHIKV to replicate in targeted cell types it is essential for the virus to enter these cells. In this review, we summarize our current understanding of the versatile and promiscuous steps in CHIKV binding and entry into human and mosquito host cells. We describe the different entry pathways, receptors, and attachment factors so far described for CHIKV and other mosquito-borne alphaviruses and discuss them in the context of tissue tropism and potential therapeutic targeting. In the replication cycle of alphaviruses, the entry step is the first essential step to allow efficient production of new viral particles. The main entry pathway of alphaviruses is clathrin-mediated endocytosis. Fusion requires a low pH and takes place in the endosome. However, other, clathrin-independent, pathways that facilitate alphavirus entry have also been described. Some host factors that can mediate alphavirus entry have been identified but entry in the absence of these host factors is still possible. Alphavirus entry appears to be a promiscuous process following multiple routes where various pathways, attachment factors, and receptors can be used, assuring a broad tissue tropism. [ABSTRACT FROM AUTHOR]

The presence of a structural recognition motif for the nucleoside P2 transporter in a library of pyrimidine and triazine non-nucleoside HIV-1 reverse transcriptase inhibitors, prompted for the evaluation of antitrypanosomal activity. It was demonstrated that the structure–activity relationship for anti-HIV and antitrypanosomal activity was different. Optimization in the diaryl triazine series led to 6-(mesityloxy)- N 2 -phenyl-1,3,5-triazine-2,4-diamine ( 69 ), a compound with potent in vitro and moderate in vivo antitrypanosomal activity. [ABSTRACT FROM AUTHOR]

Abstract: This letter reports the synthesis and structure–activity relationship (SAR) study of a series of triazine dimers as novel antiviral agents. These compounds were obtained through a bivalent ligand approach in which two triazine moieties are covalently connected by suitable linkers. Several compounds showed submicromolar activity against wild-type HIV-1 and moderate activity against single mutant strains. [Copyright &y& Elsevier]

Abstract: In this Letter, we report on diarylpyridinone, diarylpyridazinone and diarylphthalazinone analogs as potential inhibitors of HIV-1 nonnucleoside reverse transcriptase (NNRTIs). The most promising compounds in these series are three diarylpyridazinones 25a, 25l and 25n which demonstrated submicromolar activity against wild-type HIV-1 and moderate activity against the single mutant strain Ba-L V106A. [Copyright &y& Elsevier]

Lung transplant recipients (LTRs) are susceptible to severe Coronavirus Disease 2019 (COVID-19) and had lower immune responses to primary severe acute respiratory syndrome-related to coronavirus 2 (SARS-CoV-2) vaccination as compared to the general population and to other solid organ transplant recipients. As immunity induced by booster vaccination and natural infection has increased since the beginning of the pandemic in the general population, immunity acquired by LTRs is not well documented. Humoral and cellular immunity to SARS-CoV-2 was monitored in February and May 2023 in 30 LTRs and compared to that of health care workers (HCWs) and nursing home residents (NHRs). LTRs had significantly lower levels of SARS-CoV-2 binding and neutralizing antibodies and lower interferon-gamma responses to Wuhan, Delta, and XBB1.5 variants as compared to HCWs and NHRs. Humoral immunity decreased between the 2 visits, whereas cellular immunity remained more stable. The persistent defect in SARS-CoV-2 immunity in LTRs should encourage continued monitoring and preventive measures for this vulnerable population. [ABSTRACT FROM AUTHOR]

• Spatiotemporal analysis of dengue virus (DENV) 1 and 2 in Africa. • Minimum 10 and eight introductions into Africa of DENV-1 and cosmopolitan DENV-2. • Recent introductions are circulating but also a long-established African clade. • After introduction into Africa, a limited geographical spread can be observed. • Active genomic surveillance might limit sero- and genotype spread and disease. The origin and spread of dengue virus (DENV) circulating in Africa remain poorly characterized, with African sequences representing <1% of global sequence data. Whole genome sequencing was performed on serum samples (n = 29) from an undifferentiated fever study in 2016 in the Democratic Republic of Congo (DRC), and from febrile travelers returning from Africa. The evolutionary history of the newly acquired African DENV-1 (n = 1) and cosmopolitan genotype DENV-2 (n = 18) genomes was reconstructed using a phylogeographic, time-scaled Bayesian analysis on a curated DENV panel including all known African sequences. A minimum of 10 and eight introductions could be identified into Africa for DENV-1 and cosmopolitan DENV-2, respectively, almost all originating from Asia. Three introductions were previously unknown. The currently circulating virus comprises mainly the recently introduced clades and one long-established African clade. Robust geographical clustering suggests limited spread of DENV after each introduction. Our data identified the DRC as the source of the 2018 Angolan DENV-2 epidemic, and similarly, the 2013 Angolan DENV-1 outbreak as the origin of our DRC study. Active genomic surveillance of DENV in Africa at the portals of entry might help early outbreak response and limit sero- and genotype spread and human disease burden. [ABSTRACT FROM AUTHOR]

To overcome supply issues of COVID-19 vaccines, this partially single blind, multi–centric, vaccine trial aimed to evaluate humoral immunogenicity using lower vaccine doses, intradermal vaccination, and heterologous vaccine schedules. Also, the immunity after a booster vaccination was assessed. 566 COVID-19-naïve healthy adults were randomized to 1 of 8 treatment arms consisting of combinations of BNT162b2, mRNA-1273, and ChAdOx1-S. Anti-Receptor-Binding Domain immunoglobulin G (RBD IgG) titers, neutralizing antibody titres, and avidity of the anti-RBD IgGs was assessed up to 1 year after study start. Prolonging the interval between vaccinations from 28 to 84 days and the use of a heterologous BNT162b2 + mRNA-1273 vaccination schedule led to a non-inferior immune response, compared to the reference schedule. A low dose of mRNA-1273 was sufficient to induce non-inferior immunity. Non-inferiority could not be demonstrated for intradermal vaccination. For all adapted vaccination schedules, anti-RBD IgG titres measured after a first booster vaccination were non-inferior to their reference schedule. This study suggests that reference vaccine schedules can be adapted without jeopardizing the development of an adequate immune response. Immunity after a booster vaccination did not depend on the dose or brand of the booster vaccine, which is relevant for future booster campaigns. The trial is registered in the European Union Clinical Trials Register (number 2021–001993-52) and on clinicaltrials.gov (NCT06189040). [ABSTRACT FROM AUTHOR]

Background: SARS-CoV-2 vaccination is strongly recommended in kidney transplant recipients (KTR) and dialysis patients. Whether these vaccinations may trigger alloantibodies, is still debated. Methods: In the current study we evaluated the effect of SARS-CoV-2 mRNA vaccines on anti-Human Leukocyte Antigen (HLA) and 60 anti-non-HLA antibody profiles in clinically stable KTR and dialysis patients. In total, we included 28 KTR, 30 patients on haemodialysis, 25 patients on peritoneal dialysis and 31 controls with a positive seroresponse 16-21 days after the first dose of either the SARS-CoV-2 mRNA BNT162b2 or mRNA-1273 vaccine. Both anti-HLA and anti-non-HLA antibodies were determined prior to vaccination and 21 to 35 days after the second vaccine dose. Results: Overall, the proportion of patients with detectable anti-HLA antibodies was similar before and after vaccination (class I 14% vs. 16%, p = 0.48; class II 25% before and after vaccination). After vaccination, there was no pattern in 1) additionally detected anti-HLA antibodies, or 2) the levels of pre-existing ones. Additional anti-non-HLA antibodies were detected in 30% of the patients, ranging from 1 to 5 new anti-non-HLA antibodies per patient. However, the clinical significance of anti-non-HLA antibodies is still a matter of debate. To date, only a significant association has been found for anti-non-HLA ARHGDIB antibodies and long-term kidney graft loss. No additionally developed anti-ARHGDIB antibodies or elevated level of existing anti-ARHGDIB antibodies was observed. Conclusion: The current data indicate that SARS-CoV-2 mRNA vaccination does not induce anti-HLA or anti-non-HLA antibodies, corroborating the importance of vaccinating KTR and dialysis patients. [ABSTRACT FROM AUTHOR]

• Integrated data sources reveal insights into 2022s mpox epidemic decline. • Mpox first affected individuals with high-, then with lower-risk sexual behavior. • Seemingly consistent sexual behavior in population at risk throughout mpox outbreak. • Vaccination timing and seroprevalence suggest minor role of population immunity. • Continued vigilance and surveillance remain essential to identify new outbreaks. The 2022 mpox epidemic reached a peak in Belgium and the rest of Europe in July 2022, after which it unexpectedly subsided. This study investigates epidemiological, behavioral, and immunological factors behind the waning of the epidemic in Belgium. We investigated temporal evolutions in the characteristics and behavior of mpox patients using national surveillance data and data from a prospective registry of mpox patients in the Institute of Tropical Medicine (Antwerp). We studied behavioral changes in the population at risk using a survey among HIV-preexposure prophylaxis (PrEP) users. We determined the seroprevalence of anti-orthopoxvirus antibodies among HIV-PrEP users across four-time points in 2022. Mpox patients diagnosed at the end of the epidemic had less sexual risk behavior compared to those diagnosed earlier: they engaged less in sex at mass events, had fewer sexual partners, and were less likely to belong to the sexual network's central group. Among HIV-PrEP users there were no notable changes in sexual behavior. Anti-orthopoxvirus seroprevalence did not notably increase before the start of national vaccination campaigns. The observed changes in group immunity and behavior in the population at greater risk of exposure to mpox seem unable to explain the waning of the mpox epidemic. A change in the profile of mpox patients might have contributed to the decline in cases. [ABSTRACT FROM AUTHOR]

Background: The Democratic Republic of the Congo has confronted 13 outbreaks of Ebola virus disease since 1976. Rapid diagnostic tests (RDTs) detecting viral antigens have been developed to circumvent difficulties encountered with RT-PCR for diagnosis in remote low-resource settings, but there is still uncertainty about their performance characteristics and usability during outbreaks. We aimed to assess the field performance of three antigen detection RDTs compared with the gold-standard Cepheid GeneXpert Ebola assay results.Methods: We conducted a retrospective, multicentre observational study using complete and de-identified databases of five mobile laboratories (managed by the Institut National de Recherche Biomédicale) to assess the performance of three Ebola virus disease RDTs (QuickNavi-Ebola, OraQuick Ebola Rapid Antigen Test, and Coris EBOLA Ag K-SeT rapid test) run on blood samples of patients with suspected Ebola virus disease in direct comparison with the Cepheid GeneXpert Ebola assay reference test during the 2018-20 outbreak in the eastern Democratic Republic of the Congo. We estimated the sensitivity and specificity of each test through generalised linear mixed models against the GeneXpert Ebola assay reference test and corrected for cycle threshold value and random site effects.Findings: 719 (7·9%) of 9157 samples had a positive GeneXpert Ebola assay result. The QuickNavi-Ebola RDT had a sensitivity of 87·4% (95% CI 63·6-96·8) around the mean cycle threshold value and a specificity of 99·6% (99·3-99·8). The OraQuick Ebola Rapid Antigen Test had a sensitivity of 57·4% (95% CI 38·8-75·8) and specificity of 98·3% (97·5-99·0), and the Coris EBOLA Ag K-SeT rapid test had a sensitivity of 38·9% (23·0-63·6) against the GeneXpert Ebola assay reference and specificity of 97·4% (85·3-99·6). The QuickNavi-Ebola RDT showed a robust performance with good sensitivity, particularly with increasing viral loads (ie, low cycle threshold values), and specificity.Interpretation: The three RDTs evaluated did not achieve the desired sensitivity and specificity of the WHO target product profile. Although the RDTs cannot triage and rule out Ebola virus infection among clinical suspects, they can still help to sort people with suspected Ebola virus disease into high-risk and low-risk groups while waiting for GeneXpert Ebola assay reference testing.Funding: None.Translation: For the French translation of the abstract see Supplementary Materials section. [ABSTRACT FROM AUTHOR]

Infections with arthropod-borne viruses are increasing globally as a result of climate and demographic changes, global dispersion of insect vectors, and increased air travel. The similar symptomatology of arboviral diseases and the cocirculation of different arboviruses in Africa, Asia, and South America complicate diagnosis. Despite the high sensitivity and specificity of molecular diagnostic tests, their utility is limited to the short viremic phase of arbovirus infections, and therefore the diagnosis of infection is frequently missed in clinical practice. Conversely, the duration of antibody responses provides a wider window of opportunity, making diagnosis more dependent on IgM/IgG detection. This review discusses the issues underlying the low specificity of antibody-detection assays, and addresses the challenges and strategies for discovering more specific biomarkers to enable a more accurate diagnosis. Highly specific diagnostic tests for arbovirus infection are required that are easy to use, affordable, and applicable in low- and middle-income countries. When the acute phase is missed, diagnosis relies on detection of IgM, IgG, or both, in convalescent serum. This allows the diagnosis of past arboviral infections. Antigen-detection tests are useful as a first-line test in endemic areas. Currently, only a handful of validated commercial antigen-detection tests are available for dengue virus (DENV) and Zika virus (ZIKA), which suffer from sensitivity issues. Additionally, antibody-detection methods lack good specificity because flaviviruses and other cocirculating pathogens share a degree of sequence and structural similarity, resulting in immunological cross-reactivity. Being able to distinguish between type-specific and cross-reactive antibodies is important for clinical diagnosis (e.g., increased risk for severe disease upon secondary heterologous DENV infection, neurological complications, Zika-related developmental disorders), for serostatus determination related to DENV vaccination, and – once available – for specific antiviral treatment. [ABSTRACT FROM AUTHOR]

• We report on a new dengue virus variant circulating in the Peruvian Amazon. • Genotypic characterization of dengue virus is useful to monitor the distribution of circulating genotypes in endemic areas. • Monitoring the introduction of new dengue virus strains helps to understand key drivers of dengue viral dynamics and epidemic patterns. We sequenced the envelope gene of dengue virus serotype 2 (DENV-2-E) in samples from an outbreak reported in 2018, in Yurimaguas, Peru. The strain belongs to lineage 2 of the American/Asian genotype. We report a variant with two novel mutations (I379T and V484I) located in domain III of DENV2-E. [ABSTRACT FROM AUTHOR]

• Diagnostic sensitivity of alternative sampling specimens for MPXV PCR are described. • Saliva contains higher MPXV viral load than oropharyngeal swabs. • Plasma and serum samples do not contain high MPXV viral load. • Saliva MPXV viral load warrants elucidating its role in mpox transmission. Mpox (formerly monkeypox) is a viral disease caused by the mpox virus (MPXV), endemic in Central and West Africa and currently causing a global outbreak of international concern. Much remains unknown about sample types most suited for mpox laboratory diagnosis. While it is established that high viral loads can be found in active skin lesions (currently the recommended mpox laboratory confirmation specimen type), WHO mpox testing guidelines encourage the use of oropharyngeal swabs as an additional sample type for mpox diagnosis and suggest investigating the value of other specimens like blood samples. In this study, we verified the value of select alternative specimen types for mpox laboratory confirmation. We included 25 patients with MPXV-confirmed skin lesions to compare diagnostic sensitivity of MPXV PCR testing on EDTA plasma and two upper respiratory specimens: oropharyngeal swabs and saliva. In our patient cohort with MPXV-confirmed skin lesions, diagnostic sensitivity of MPXV PCR was 80% in EDTA plasma, 64% in oropharyngeal swabs, and 88% in saliva. MPXV viral loads were significantly higher in saliva compared to oropharyngeal swabs and EDTA plasma. The WHO recommendation to collect oropharyngeal swabs as an additional specimen for mpox diagnosis might need to be revised to include saliva wherever feasible. We suggest investigating saliva as a diagnostic specimen in the absence of active skin lesions or during the phase preceding skin manifestations. Moreover, the relatively high MPXV DNA content of saliva warrants elucidating its potential role in disease transmission. [ABSTRACT FROM AUTHOR]

We report the first laboratory-confirmed Zika virus (ZIKV) infection in a Belgian traveler after a three week holiday in Guatemala, December 2015. This case along with other imported cases into Europe emphases once again the need for accurate diagnostic tools for this rapidly emerging virus. The challenge is to diagnose patients in the acute phase, which appears short, as serological testing is complicated by cross-reactivity, vaccination status and scarce availability of specific ZIKV tests. [ABSTRACT FROM AUTHOR]

● Sciensano and its department Quality of Laboratories routinely organizes external quality assessment as a part of the mandatory external quality assessment program for SARS-CoV-2 serology in Belgium. ● Sample IS/17575 generated highly discordant results. ● The importance of the choice of antigenic composition. ● Antibody response against the N 184 protein appears to wane post-infection. ● Difference in sensitivity of RBD 192 vs S1 vs S protein based immunoassays. We report on sample IS/17575 since it generated highly divergent results in the Belgian SARS-CoV-2 serology external quality assessment scheme. Sample IS/17575 was serum originating from a 30 years old male patient. 124 diagnostic laboratories analysed this sample. A total of 168 results was returned (including 5 doubles). Overall, 38 were positive. All tests against S1 were positive except the Euroimmun IgG ELISA and the Ortho clinical Diagnostics VITROS IgG CLIA. All tests against S1/S2 (Liaison, Diasorin) resulted in a signal above cutoff. Assays against RBD, mostly generate a negative result. An exception are the Wantai SARS-CoV-2 ELISA's. All tests targeting N protein were negative. The survey shows, when >6 months post-infection, assays targeting at least S1, and preferably S1 combined with S2, are the most sensitive. This finding accentuates the necessity of external quality assessment schedules and importance of antigenic composition of serologic SARS-CoV-2 assays. [ABSTRACT FROM AUTHOR]

Chikungunya virus (CHIKV), a positive-sense, single-stranded RNA virus in the family Togaviridae, is transmitted by Aedes mosquitoes. Of three known CHIKV genotypes, the Asian genotype was introduced into the Caribbean islands and rapidly spread throughout Central and South Americas. We previously found patients with symptoms compatible with chikungunya fever in 2014–2015 in Aruba, a Caribbean island of 180 km2. We here describe the full genome sequences of eight CHIKV strains isolated from patient sera of the Aruban outbreak. Phylogenetic analysis revealed that two closely related but distinct lineages of Asian-genotype CHIKV circulated simultaneously during the epidemic in 2014–2015. These results suggested that CHIKV was introduced into Aruba more than once in a short period, reflecting the importance of Aruba as a travel hub within the region. • During the 2014-2015 Chikungunya virus (CHIKV) outbreak in a Caribbean island Aruba, we obtained 8 CHIKV isolates. • Phylogenetic analysis of these isolates revealed that two distinct lineages of Asian-genotype CHIKV cocirculated. • We confirmed Caribbean outbreak lineage specific duplication in the 3' untranslated region of these isolates. [ABSTRACT FROM AUTHOR]

Quantitative measurement of viral load is an important parameter in the management of filovirus disease outbreaks because viral load correlates with severity of disease, survival, and infectivity. During the ongoing Ebola virus disease outbreak in parts of Western Africa, most assays used in the detection of Ebola virus disease by more than 44 diagnostic laboratories yielded qualitative results. Regulatory hurdles involved in validating quantitative assays and the urgent need for a rapid Ebola virus disease diagnosis precluded development of validated quantitative assays during the outbreak. Because of sparse quantitative data obtained from these outbreaks, opportunities for study of correlations between patient outcome, changes in viral load during the course of an outbreak, disease course in asymptomatic individuals, and the potential for virus transmission between infected patients and contacts have been limited. We strongly urge the continued development of quantitative viral load assays to carefully evaluate these parameters in future outbreaks of filovirus disease. [ABSTRACT FROM AUTHOR]