Virus Concentration Research Articles

Detection and quantification of human immunodeficiency virus-1 (HIV-1) total nucleic acids in wastewater settled solids from two California communities

ABSTRACT Wastewater surveillance for infectious agents has proved useful in identifying the circulation of viruses within populations. We investigated the presence and concentration of human immunodeficiency virus (HIV)-1 total nucleic acids (including both viral RNA and proviral DNA) in wastewater solids. We retrospectively measured HIV-1 nucleic acids in two samples per week for 26 months at two wastewater treatment plants serving populations with different prevalences of HIV infections in San Francisco and Santa Clara County, California, USA. We detected HIV nucleic acids in a majority of samples with concentrations ranging from non-detect to 3.9 × 10 5 cp/g ( N = 459 samples total). Concentrations of HIV-1 were significantly higher in samples from the wastewater treatment plant serving a population with a higher prevalence of people living with HIV than in the plant serving a population with a lower prevalence. The HIV-1 nucleic acids amplified were primarily DNA and thus represented proviral DNA shedding into wastewater. Additionally, we found that HIV-1 nucleic acid concentrations in wastewater solids were orders of magnitude higher than those in liquid wastewater indicating that the HIV-1 target preferentially sorbs to solids. Whether concentrations of HIV-1 in wastewater solids can be used to identify the number of incident cases remains unknown. Additional work on HIV-1 shedding from individuals with viremia and people living with HIV is needed to translate wastewater measurements into quantitative information on infections. Additional work may also be needed to document non-human sources of HIV-1 nucleic acids in wastewater. IMPORTANCE Human immunodeficiency virus (HIV)-1 has infected nearly 100 million people since it emerged in the 1980s. Antiretroviral therapy prevents transmission of HIV and also allows infected individuals to live healthy lives with normal life expectancy. Consequently, identifying unrecognized cases of HIV is of paramount importance. Since wastewater represents a composite biological sample from a community, it may provide valuable information on HIV-1 prevalence that can be used to inform HIV testing and outreach.

Comparative analysis of whole-genome sequences of African swine fever virus (Asfarviridae: Asfivirus) isolates сollected on the territory of the left bank of the Dnieper River in 2023

Introduction. The lack of data on the whole-genome sequences of African swine fever virus (ASFV) variants circulating on the territory of the left bank of the Dnieper River complicates the understanding of the molecular evolution of the virus and the character of the epidemic process development in Russia and Ukraine. Understanding the genetic divergence and phylogenetic relatedness of isolates can largely adjust the strategy of general and specific prevention of the disease. The aim of the study – search and description of unique mutations (deletions/insertions/substitutions) in isolates collected from domestic pigs in Donetsk, Luhansk and Zaporozhye regions in 2023; determination of relatedness and level of homology with reference strains of ASFV genotype II; sub-genotyping and clustering of isolates based on whole-genome analysis. Materials and methods. The samples used were a culture suspension of porcine bone marrow (PBM) cells containing ASFV isolates obtained from pathologic material from domestic pig carcasses. Genomic DNA was prepared by purification and concentration of virus followed by phenol-chloroform extraction of total nucleic acid. The high-throughput sequencing process was performed using MGI technology. Consensus sequences were assembled by mapping reads to the reference genome of strain Georgia 2007/1. Results. All isolates are assigned to genotype II, have a monophyletic origin, are phylogenetically close to the clusters «Europe» (4/5) and «Bryansk 2021» (1/5), and are divergent from the original parental genetic variants that make up the enlarged clades. In addition, numerous substitutions in the loci of the multigene family MGF 110, 505, and 360, encoding virulence proteins, were detected in 4 isolates from Donetsk and Zaporozhye regions. Conclusion. The phylogeny of the genotype II ASFV, which originated from the reference strain Georgia 2007/1, is shown to be sufficient for isolate differentiation. The presented data are of theoretical and practical importance for domestic and international ASFV surveillance.

Transmission of Cryphonectria Hypovirus 1 (CHV1) to Cryphonectria radicalis and In Vitro and In Vivo Testing of Its Potential for Use as Biocontrol Against C. parasitica

Cryphonectria hypovirus 1 (CHV1) is successful in controlling Cryphonectria parasitica, the causal agent of chestnut blight, but little is known regarding its transmission to other fungi, for example the European Cryphonectria radicalis. In this study, CHV1 was transmitted (circa 200,000–800,000 copies/microliter) to seven C. radicalis isolates from infected C. parasitica. Reverse transmission to virus-free C. parasitica (European 74 testers collection) was achieved, although it was less successful (250–55,000 copies/µL) and was dependent on the vegetative compatibility (VC) group. In C. radicalis, the virus infection led to colony colour change from pink to white and smaller colonies, dependent on the virus concentration. The virus was concentrated in the colony edges, and vertically transmitted to 77% of conidia. However, several in vitro experiments demonstrated that C. radicalis was always outcompeted by the blight fungus, only suppressing the pathogen between its 25–50% inoculum level. It presented good secondary capture only when acting as a pioneer. Two types of in planta assays (individual and challenge inoculations) were undertaken. Cryphonectria radicalis behaved as a saprotroph, while chestnut blight fungus behaved as an aggressive pathogen, and lesions after treatment with C. radicalis were no smaller in general, only when using cut branches. Overall, the results showed that infected C. radicalis was unable to control cankers.

From Viral Content to Real-Life Cuisine and Beyond: Examining Teenagers' Interactions with TikTok Food Videos and the Influence on their Food Practices

TikTok has gained immense popularity among teenagers, offering access to numerous user-generated content. Notably, food videos have emerged as a prominent theme on this short-form video social platform. The casual and enjoyable nature of short food videos on TikTok belies their potential influence on one of teenagers' most immediate and regular health practices--eating. Understanding how teenagers interact with these videos, their subsequent actions, and the resulting impact on their food practices and eating habits have the potential to provide insight into their broader lifestyle choices and their interactions within their social circles, including parents, friends, and other people online. By examining how teenagers use TikTok food videos online and offline, we gain a deeper understanding of the intricate relationship between social media, teenage lifestyle, and social dynamics surrounding food practices. In this research, we conducted 15 semi-structured interviews with teenagers aged 13 to 19, investigating their consumption of TikTok food videos and the actions inspired by them. By examining the multifaceted influence of TikTok food videos from a temporal perspective, this study contributes to the reflections of teens' use of TikTok food videos and their inspired food practices in the short and long term, online and offline. We propose design and theoretical implications to support teenagers' health. These insights have the potential to extend to various contexts, helping educators, policymakers, and designers in fostering healthy lifestyles among teenagers.

Double infection of Nicotiana benthamiana with AMV and WCMV increases both virus concentrations and synergistically changes both host organelle ultrastructure and chlorophyll content

To clarify the synergistic pathogenic mechanism of Nicotiana benthamiana double infection with alfalfa mosaic virus (AMV) and white clover mosaic virus (WCMV), AMV and WCMV co-inoculation of N. benthamiana as treatment and single inoculation of AMV or WCMV and phosphate buffer solution (pH 7.0, PBS) as control, respectively. The concentrations and the relative expression of AMV and WCMV coat proteins were determined by a double antibody sandwich enzyme-linked immune sorbent assay (DAS-ELISA) and real-time fluorescence quantitative PCR (RT-qPCR) in a double infection of N. benthamiana with AMV and WCMV. Meanwhile, virion morphology, ultrastructure morphology, and chlorophyll content were observed and determined by electron microscopy. The results showed that the diseased symptoms were more serious, and virus concentration and relative expression of AMV and WCMV coat proteins were also higher in N. benthamiana double infection with AMV and WCMV than in AMV or WCMV single infection. The main symptoms manifested as severe mottle mosaic, shrinkage, and chlorosis. The concentrations of AMV and WCMV were 182.23pg/mL and 148.77pg/mL of double infection with AMV and WCMV, which were 1.75-fold and 1.62-fold than AMV and WCMV single infection, respectively. The relative expression of AMV and WCMV coat proteins was 4.25-fold and 2.50-fold than the single virus infection, respectively. Electron microscopy also observed that chloroplast malformation, cell membrane deformation, contents dissolution, grana lamella disorder, fat granules increased and enlarged, starch granules enlarged, and mitochondria were seriously malformed in a double infection of N. benthamiana with AMV and WCMV. The chlorophyll content was significantly lower for double infection with AMV and WCMV than for AMV or WCMV single-infected and CK, reduced by 31.52%, 22.83%, and 76.09%, respectively. This is the first report of a double infection of N. benthamiana with AMV and WCMV that increases both virus concentrations and synergistically changes both host organelle ultrastructure and chlorophyll content.

Numerical simulation of the dual-loop different velocity air curtain system in the operating room

The problem of cross-infection between patients and surgeons in the operating room had been widely concerned by the international community. The purpose of this study had been to explore the potential of a Dual-loop Different Velocity Air Curtain System (DDVACS) being applied in the operating room to reduce cross-infection. In this study, the performance of the system was studied using the computational fluid dynamics (CFD) numerical simulation method, with the operating conditions of different air supply velocities inside and outside being set. The RNG k-ε model, Discrete Phase Model (DPM), and Species Transport model were used to simulate the air distribution, particle diffusion, and virus diffusion. The main research results showed that, when the DDVACS was operated at an inner air supply velocity of 0.24 m/s and an outer air supply velocity of 0.45 m/s, the virus concentration in the surgical area had decreased by 89%, the particulate matter concentration in the surgical area had decreased by 78.08%, but the deposition of particulate matter had increased by 21.62%. This study concluded that DDVACS had been found to effectively reduce the concentration of cross-infection. Finally, the velocity difference between the inner and outer sides was controlled within 2 times. This setting had been arranged to make the DDVACS work better. This study's contribution was to further verify the application of the DDVACS in operating rooms, providing a new perspective and empirical support for related fields and possessing certain practical application value.

Antioxidant anthocyanin synergistic immune enhancer nanopeptide C-I20 remarkably enhances the protective effect of largemouth bass against largemouth bass ranavirus

Largemouth bass ranavirus (LMBV) infection results in huge economic losses in largemouth bass (Micropterus salmoides) industry. Nanopeptide C-I20 and anthocyanins have a positive effect on promoting immune responses and antioxidant mechanisms in several aquatic organisms, and are therefore used to inhibit LMBV infection. In this study, we developed an LMBV immersion challenge model using three different viral concentrations (1×104 copies/mL, 1×105 copies/mL, and 1×106 copies/mL) to infect largemouth bass, and LMBV-MCP mRNA expression was detected in infected fish. Following infection, the fish exhibited severe external ulceration, redness swelling, and darkening of the skin. Histopathological examination revealed significant necrosis and inflammation in muscle tissue, epithelial cell shedding in renal tubules, macrophage aggregation centers and cellular vacuolization in spleen and head kidney, and cellular hypertrophy in liver. To mitigate LMBV infection, we explored the protective effects of a combined treatment strategy involving C-I20 and anthocyanin. Overall, the combination of anthocyanin and C-I20 demonstrated the highest protective efficacy, significantly reducing viral loads in muscle, liver, spleen, and head kidney. Moreover, this treatment regimen enhanced antioxidant enzyme activities (T-AOC, TSOD, GSH-Px, CAT) and modulated important immune genes (IL-1, IL-8, TNF-α, IL-10, Mx, and IgM) expression. In conclusion, the synergistic application of anthocyanin and C-I20 demonstrates significant efficacy in mitigating LMBV infection. This research introduces a novel and promising approach to managing infectious diseases in aquaculture settings.

Influencing Factors Analysis of Infectious SARS-CoV-2 Aerosols Sampling

Assessing the potential infectivity of airborne viruses is critical for evaluating the risk of their transmission through air. This study investigated the factors influencing the collection of infectious SARS-CoV-2 aerosols using three common aerosol samplers: the impactor sampler AGI-30, the SKC Biosampler, and a cyclone sampler WA-400III. It was found that the sampling process over time significantly impacted the infectivity of SARS-CoV-2 captured in the sampler, and the infectivity loss of different SARS-CoV-2 variants in the process varied. Additionally, adding newborn calf serum in the collection suspension could effectively preserve viral infectivity in the sampler. Further tests conducted under various ventilation occasions indicated that ventilation can reduce the virus concentration in the environment and rapidly clear viral particles after aerosol generation. Although the flow rate of WA-400III is higher, it only could collect higher concentrations of viral RNA instead of live viruses than AGI-30 or SKC Biosampler when aerosol was generated constantly in the environment. After aerosol generation stopped, the WA-400III collected more infectious viruses and viral RNA. This study highlights the impact of the sampling process on captured virus should be assessed and protective agent is needed to preserve viral viability. And it is crucial to select appropriate sampler based on environmental characteristics and research objectives for obtaining optimal results. The AGI-30 and SKC Biosampler are preferable for collecting infectious viruses in environments with high aerosol concentrations, while the cyclone sampler WA-400III is more effective for collecting viral nucleic acids and enriching virus samples in confined spaces with lower viral loads.

Development of a concentration method for simple and sensitive detection of SARS-CoV-2 in saliva using a magnetic nanoparticle kit

BackgroundWhen diagnosing viral infections in humans and animals, the presence of virus in a sample in trace amounts that are below the analytical sensitivity of the detection system may cause false negative results and inaccurate diagnosis. We previously reported the development of a simple virion concentration technique using 12ml large-volume samples that can dramatically improve diagnostic sensitivity by increasing analytical sensitivity by 100-fold over conventional methods. The present study was conducted to further improve the simplicity and versatility of this method. We constructed a simple and highly sensitive method for the detection of SARS-CoV-2 in human saliva after concentration using a magnetic nanoparticle conjugated with polyethylene glycol (PEG). ResultsPerformance of the method was evaluated by comparing a combination of automated nucleic acid extraction and RT-qPCR or triplex RT-LAM detection in a spiked sample of 20ml saliva collected from healthy humans. The method theoretically achieved 300-fold concentration of spiked SARS-CoV-2 in saliva, enabling 10- to 1000-fold higher analytical sensitivity for detection compared to conventional RNA extraction methods. ConclusionsThis newly developed method allows for easy and reliable concentration of the virion in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. Further, the method allows for easy and reliable enrichment of the virus in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. This method is easily used for highly sensitive virus detection from a variety of human oral fluid samples and may also be applied to rapid and labor-saving screening tests by pooling a large number of samples.

An Exploratory Study of Fact-Checking Practices in Conflict and Authoritarian Contexts

This study explores fact-checking practices in Ethiopia and Mali in times of conflict and in a context marked by increasing restrictions to press freedom. The objective is to understand how, in this hostile environment, fact-checkers in these two countries manage to carry out their activities. Our findings reveal that fact-checkers are often victims of online bullying and harassment and fear reprisal from governments. This pushes them to self-censor, avoiding working on sensitive topics, such as military issues in Mali. In addition, fact-checking organizations in both countries highlight the difficulty of accessing reliable sources. Consequently, they focus more on debunking viral social media content, thus effectively becoming content moderators who have turned away from the mission of holding leaders accountable, one of the primary functions of fact-checking. Regarding their role conception, fact-checkers in Ethiopia and Mali see themselves more as guides helping navigate the information disorder than “guardians of truth” or “truth keepers.”

Droplet Digital PCR Enhances Sensitivity of Canine Distemper Virus Detection

Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR’s superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis.

Steganographic Metadata Embedding to Trace the Footprint of WhatsApp Images

This research paper proposes a method to trace the footprint of senders while forwading of images on WhatsApp by embedding metadata (such as mobile numbers) using Least Significant Bit (LSB) steganography. The technique ensures that the hidden information is imperceptible to users and remains intact even after WhatsApp’s image compression. The method allows for tracking the digital footprint of images, providing traceability to combat misinformation, piracy, and privacy concerns. The system is robust, enabling metadata recovery after multiple shares, with applications in digital forensics, media integrity, and monitoring the spread of viral content

Investigation of the Impact of New Media on Movie Advertising

As the landscape of mass entertainment shifts towards new media—characterized by short-form videos, interactive content, and decentralized platforms—the traditional film industry is undergoing significant changes in its advertising strategies. This paper investigates the impact of new media on movie advertising by analyzing how digital channels, such as social media and streaming services, have transformed traditional promotional methods. Utilizing reliable secondary sources, this research explores various aspects of new media’s influence on film marketing, focusing on case studies like “Avengers: Endgame” and “Spider-Man: No Way Home” to highlight effective strategies. The findings reveal that new media offers unprecedented opportunities for audience engagement through dynamic and interactive content, surpassing the effectiveness of traditional methods. Social media campaigns, marked by high levels of engagement, viral content, and influencer collaborations, have proven particularly successful in driving film visibility and box office success. The research concludes that future studies should incorporate emerging platforms like TikTok, which present novel ways to connect with younger demographics and create shareable content. By understanding the evolving trends and integrating new media strategies, filmmakers can optimize their advertising approaches to enhance audience reach and maximize commercial success.

ATPase Valosin-Containing Protein (VCP) Is Involved During the Replication and Egress of Sialodacryoadenitis Virus (SDAV) in Neurons

Sialodacryoadenitis virus (SDAV) has been identified as the etiological agent responsible for the respiratory system and salivary gland infections in rats. The existing literature on SDAV infections is insufficient to address the topic adequately, particularly in relation to the central nervous system. In order to ascertain how SDAV gains access to neuronal cells and subsequently exits, our attention was focused on the small molecule valosin-containing protein (VCP), which is an ATPase. VCP is acknowledged for its function in the ubiquitin-mediated proteasomal degradation of proteins, including those of viral origin. To ascertain the potential influence of VCP on SDAV replication and egress, high-content screening was employed to determine the viral titer and protein content. Western blot analysis was employed to ascertain the relative expression of VCP. Real-time imaging of SDAV-infected cells and confocal imaging for qualitative morphological analysis were conducted. The Eeyarestatin I (EerI) inhibitor was employed to disrupt VCP involvement in the endoplasmic reticulum-associated protein degradation pathway (ERAD) in both pre- and post-incubation systems, with concentrations of 5 μM/mL and 25 μM/mL, respectively. We demonstrated for the first time that SDAV productively replicates in cultured primary neurons. VCP expression is markedly elevated during SDAV infection. The application of 5 μM/mL EerI in the post-treatment system yielded a statistically significant inhibition of the SDAV yield. It is likely that this modulates the efficacy of virion assembly by arresting viral proteins in the submembrane area.

Wastewater Surveillance of SARS-CoV-2 in Slovenia: Key Public Health Tool in Endemic Time of COVID-19

With the reclassification of COVID-19 as an endemic disease and the relaxation of measures, Slovenia needed a complementary system for monitoring SARS-CoV-2 infections. This article provides an overview of the epidemiological situation of SARS-CoV-2 in Slovenia using a wastewater surveillance system, demonstrating its usefulness as a complementary tool in epidemiological surveillance. This study found that estimated SARS-CoV-2 infections in Slovenia peaked in September 2022 and showed a declining trend with subsequent lower peaks in March–April and December 2023, mirroring the trends observed from clinical data. Based on both surveillance systems, the most prevalent variant in 2022 was BA.5. By 2023, BQ.1 and other Omicron variants increased in prevalence. By the end of 2023, XBB sublineages and the BA.2.86 variant had become predominant, demonstrating consistent dynamic shifts in variant distribution across both monitoring methods. This study found that wastewater surveillance at wastewater treatment plants in Slovenia effectively tracked SARS-CoV-2 infection trends, showing a moderate to strong correlation with clinical data and providing early indications of changes in infection trends and variant emergence. Despite limitations during periods of low virus concentration, the system proved significant in providing early warnings of infection trends and variant emergence, thus enhancing public health response capabilities.

Air cleaner prototype: Reduction of airborne viruses and effects of UV-C irradiation on virus concentration and RNA copy numbers considering modeled residence times and doses

Air cleaner prototype: Reduction of airborne viruses and effects of UV-C irradiation on virus concentration and RNA copy numbers considering modeled residence times and doses

Wastewater-Based Epidemiology of Influenza Viruses: a systematic review

Abstract Background Influenza viruses (IVs) are responsible for annual epidemic outbreaks during the winter months. Beyond conventional surveillance systems, the circulation of these pathogens can be monitored using Wastewater-Based Epidemiology (WBE), an epidemiological tool that describes the health status of an entire community by analyzing the presence of pathogens within wastewater. Systematically reviewing existing experiences in this emerging field is crucial to highlight the best practices to establish a WBE surveillance system providing useful information for public health actions. Methods The study protocol for this systematic review has been registered on the PROSPERO website (registration ID: CRD42024532435). Three academic databases (PubMed, Web of Science and Scopus) were accessed to collect studies examining the presence of IVs in wastewater. Studies included were those reporting quantitative measures of viral concentration or detailing molecular/phylogenetic characterization of IVs in wastewater. Results Databases were searched on July 8th, 2024. After de-duplication and screening of the 400 remaining records, 42 were included in the review. IVs detection in wastewater was reported in all papers except from one, although with high variability in terms of positivity titers and concentration levels. More than half of the collected papers (52.4%) evaluated association between environmental viral concentration and clinical data, generally showing concordance between the two aspects. Moreover, 28.6% of papers described subtyping of IVs in wastewater through different methods (specific PCR assays being the most used). Conclusions WBE is a promising approach for surveilling viral circulation, and IVs represent a high value target for implementing this innovative system. Systematically reviewing previous experiences can be useful to obtain information for the establishment of a novel environmental surveillance system of Influenza Viruses Key messages • Nucleic acids belonging to Influenza Virus are detectable in wastewater, even though we report high levels of variability in sampling techniques, laboratory methods and data management systems. • Association between environmental data and number of clinical cases is reported to be generally strong in collected papers; subtyping of IVs present in wastewater is also feasible.

Etyczne projektowanie w dobie generatywnej sztucznej inteligencji

Implementation of AI solutions is unavoidable on the markets, where the combination of time deficits and the need for constant creation of new solutions is especially demanding, that is i.a. in creative industries. Like a lens, generative AI concentrates the majority of threats and possibilities, resulting from technological changes of the recent decade. Still, despite the constant media confusion, great number of viral content, guides, courses and codes, we have much work to do towards defining the rules and specifics of designers’ collaboration with AI, and the related ethical challenges. The AI development is ahead of legal regulations and stimulates changes, and the constant information overload does not encourage reflection on AI systems and their role in design. Becoming aware of the possibilities and limitations of generative AI, however, is the only way to its sustainable and ethical use. The first step will be to reflect on and understand the possibilities and changes AI carries in the area of creation, intellectual property, and narrowing the competency gap (the lack of competency in reference to a given human team). The second step will be the efficient identification of risks and indication of new areas of responsibility to designers, who would like to apply new technologies in their work. Keywords: design, ethics, artificial intelligence (AI), responsibility, designer’s role

Sewer system sampling for wastewater-based disease surveillance: Is the work worth it?

ABSTRACT Wastewater treatment plant (WWTP) influent sampling is commonly used in wastewater-based disease surveillance to assess the circulation of pathogens in the population aggregated in a catchment area. However, the signal can be lost within the sewer network due to adsorption, degradation, and dilution processes. The present work aimed to investigate the dynamics of SARS-CoV-2 concentration in three sub-catchments of the sewer system in the city of Hildesheim, Germany, characterised by different levels of urbanisation and presence/absence of industry, and to evaluate the benefit of sub-catchment sampling compared to WWTP influent sampling. Our study shows that sampling and analysis of virus concentrations in sub-catchments with particular settlement structures allows the identification of high concentrations of the virus at a local level in the wastewater, which are lower in samples collected at the inlet of the treatment plant covering the whole catchment. Higher virus concentrations per inhabitant were found in the sub-catchments in comparison to the inlet of the WWTP. Additionally, sewer sampling provides spatially resolved concentrations of SARS-CoV-2 in the catchment area, which is important for detecting local high incidences of COVID-19.

International external quality assessment study for detection of monkeypox virus by PCR supporting laboratory preparedness during the 2022–2023 mpox outbreak and beyond

BackgroundDiagnostic capabilities and correspondent External Quality Assessments (EQA) are key for outbreak preparedness. To support diagnostic facilities with a quality assessment of newly established monkeypox virus (MPXV) molecular diagnostic workflows, Quality Control for Molecular Diagnostics (QCMD) and the Bundeswehr Institute of Microbiology (IMB) piloted an international EQA study conducting four challenges from autumn 2022 to summer 2023 during the global mpox outbreak. ObjectivesTo assess the performance (sensitivity/specificity) of molecular assays used by diagnostic laboratories. Study designInactivated EQA panels were prepared and distributed containing seven samples of clade Ia and clade IIb MPXV strains at different viral concentrations, two specificity controls with other zoonotic orthopoxviruses (vaccinia and cowpox virus) and a negative control. Assessment was based on reported qualitative testing results. ResultsIn this outbreak-related EQA study, a total of 192 laboratories from 37 countries reported 346 qualitative datasets. Overall, core samples were correctly detected by approximately 92 % of participants in all four challenges. While sensitivity performance was acceptable in at least 90 % of datasets correctly reported even for educational MPXV-positive samples with low viral concentration [102 genome equivalents (GE)/mL], several laboratories reported the educational specificity controls as false positives or were unable to differentiate MPXV from related orthopoxviruses. ConclusionsMpox is now a globally occurring infection with a demand for quality-assured diagnostic capabilities. The newly established EQA scheme presented here, offers a multi-purpose panel for orthopoxviruses with a focus on MPXV which will continue to ensure diagnostic quality in clinical settings with up-to-date sample panels.