Universal Identification of Pathogenic Viruses by liquid chromatography coupled to tandem mass spectrometry Proteotyping.

Accurate and rapid identification of viruses is crucial for an effective medical diagnosis when dealing with infections. Conventional methods, including DNA amplification techniques or lateral-flow assays, are constrained to a specific set of targets to search for. In this study, we introduce a novel tandem mass spectrometry proteotyping-based method that offers a universal approach for the identification of pathogenic viruses and other components, eliminating the need for a priori knowledge of the sample composition. Our protocol relies on a time and cost-efficient peptide sample preparation, followed by an analysis with liquid chromatography coupled to high-resolution tandem mass spectrometry. As a proof of concept, we first assessed our method on publicly available shotgun proteomics datasets obtained from virus preparations and fecal samples of infected individuals. Successful virus identification was achieved with 53 public datasets, spanning 23 distinct viral species. Furthermore, we illustrated the method's capability to discriminate closely related viruses within the same sample, using alphaviruses as an example. The clinical applicability of our method was demonstrated by the accurate detection of the vaccinia virus in spiked saliva, a matrix of paramount clinical significance due to its non-invasive and easily obtainable nature. This innovative approach represents a significant advancement in pathogen detection and paves the way for enhanced diagnostic capabilities.

Inactivation of SARS-CoV-2 by Simulated Sunlight on Contaminated Surfaces.

We studied the stability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) under different simulated outdoor conditions by changing the temperature (20°C and 35°C), the illuminance (darkness, 10 klx, and 56 klx), and/or the cleanness of the surfaces at 50% relative humidity (RH). In darkness, the loss of viability of the virus on stainless steel is temperature dependent, but this is hidden by the effect of the sunlight from the first minutes of exposure. The virus shows a sensitivity to sunlight proportional to the illuminance intensity of the sunlight. The presence of interfering substances has a moderate effect on virus viability even with an elevated illuminance. Thus, SARS-CoV-2 is rapidly inactivated by simulated sunlight in the presence or absence of high levels of interfering substances at 20°C or 35°C and 50% relative humidity. IMPORTANCE Clinical matrix contains high levels of interfering substances. This study is the first to reveal that the presence of high levels of interfering substances had little impact on the persistence of SARS-CoV-2 on stainless steel following exposure to simulated sunlight. Thus, SARS-CoV-2 should be rapidly inactivated in outdoor environments in the presence or absence of interfering substances. Our results indicate that transmission of SARS-CoV-2 is unlikely to occur through outdoor surfaces, dependent on illuminance intensity. Moreover, most studies are interested in lineage S of SARS-CoV-2. In our experiments, we studied the stability of L-type strains, which comprise the majority of strains isolated from worldwide patients. Nevertheless, the effect of sunlight seems to be similar regardless of the strain studied, suggesting that the greater spread of certain variants is not correlated with better survival in outdoor conditions.

The variability of hepatitis B envelope is associated with HBs antigen persistence in either chronic or acute HBV genotype A infection.

BACKGROUND: More than 240 million people are chronically infected by hepatitis B virus (HBV) worldwide. Envelope proteins play a crucial role in viral cellular entry and immune recognition. The loss of HBs antigen (HBsAg) correlated with a good clinical prognosis is rarely achieved with or without treatment (3-16%). OBJECTIVES: HBV envelope variability was investigated according to HBsAg persistence. STUDY DESIGN: The cohort consisted of 15 HBV genotype A-infected patients divided into "resolvers", with HBsAg clearance, and "non-resolvers", with HBsAg persistence and in subgroups: acute (n=5, AHBV) or chronic infection (n=4, CHBV) and HBV/HIV coinfection (n=6, CHBV/HIV). HBV S and preS sequences were studied by direct and ultra-deep sequencing. Amino acid sequences were analyzed with bioinformatics for predicted antigenicity. RESULTS: In S gene, the complexity was lower in AHBV than in chronic-infected patients (p=0.046). Major mutations, detected using direct sequencing, were more frequent in AHBV developing chronicity (p=0.01) than in AHBV resolvers. In the Major Hydrophilic Region, more frequent mutations were observed in non-resolvers versus resolvers (p=0.047) and non-resolvers tended to have more haplotypes with a reduced predicted antigenicity (p=0.07). Most of the mutations in preS/S region were found rather in epitopic than in non-epitopic areas (p=0.025). Interestingly, the mutation sY161F found in 3/8 non-resolvers was associated with a decrease in predicted antigenicity (28%; AnTheProt). CONCLUSIONS: HBsAg persistence was correlated with mutations and deletions in areas playing a key role in immune recognition. These data suggest that variability in HBV envelope could favor immune escape in various clinical settings of HBV genotype A-infected patients.

Impact of deletions and mutations in Hepatitis B virus envelope proteins on serological profile and clinical evolution.

The Hepatitis B virus (HBV) envelope glycoproteins are essential for viral entry into the hepatocyte and are also targets for host immune response. The study of these proteins could allow us to highlight molecular hot points influencing HBV fitness, which would subsequently modify the clinical evolution of the disease, both under anti-viral therapy or without treatment. The present short communication underlines the importance of the high variability in HBV envelope proteins, in regard with the literature and in our hands, for HBV-infected patients either on anti-HBV treatment or not. We report mutations in antigenic areas of S protein, i.e. CD8+/CD4+ T-cell epitopes and B-cell epitopes in the major hydrophilic region (MHR), such as sI126N and sG145R possibly involved in the rare coexisting Hepatitis B surface Antigen (HBsAg)/anti-HBs serological pattern. We mostly report serial mutations in preS region including preS1 deletion (aa 1-6, 31-71, 38-73, 72-104) and preS2 deletion (aa132-141) in patients with various clinical evolutions. Some of these viral envelope mutations, due to immune selection pressure, may result in a worsening of the hepatic disease.