Microbiology spectrum最新文献

{"title":"The emergence of resistance to the antiparasitic selamectin in <i>Mycobacterium smegmatis</i> is improbable and contingent on cell wall integrity.","authors":"José Manuel Ezquerra-Aznárez, Henrich Gašparovič, Álvaro Chiner-Oms, Ainhoa Lucía, Jesús Blázquez, Iñaki Comas, Jana Korduláková, José A Aínsa, Santiago Ramón-García","doi":"10.1128/spectrum.02332-24","DOIUrl":"https://doi.org/10.1128/spectrum.02332-24","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Tuberculosis remains the deadliest infectious disease of the 21&lt;sup&gt;st&lt;/sup&gt; century. New antimicrobials are needed to improve treatment outcomes and enable therapy shortening. Drug repurposing is an alternative to the traditional drug discovery process. The avermectins are a family of macrocyclic lactones with anthelmintic activity active against &lt;i&gt;Mycobacterium tuberculosis&lt;/i&gt;. However, their mode of action in mycobacteria remains unknown. In this study, we employed traditional mutant isolation approaches using &lt;i&gt;Mycobacterium smegmatis&lt;/i&gt;, a non-pathogenic &lt;i&gt;M. tuberculosis&lt;/i&gt; surrogate. We were only able to isolate mutants with decreased susceptibility to selamectin using the ∆&lt;i&gt;nucS&lt;/i&gt; mutator &lt;i&gt;M. smegmatis&lt;/i&gt; strain. This phenotype was caused by mutations in &lt;i&gt;mps1&lt;/i&gt; and &lt;i&gt;mmpL11&lt;/i&gt;. Two of these mutants were used for a second experiment in which high-level selamectin-resistant mutants were isolated; however, specific mutations driving the phenotypic change to high-level resistance could not be identified. The susceptibility to selamectin in these mutants was restored to the basal level by subinhibitory concentrations of ethambutol. The selection of ethambutol resistance in a high-level selamectin-resistant mutant also resulted in multiple colonies becoming susceptible to selamectin again. These colonies carried mutations in &lt;i&gt;embB&lt;/i&gt;, suggesting that the integrity of the cell envelope is a prerequisite for selamectin resistance. The absence of increased susceptibility to selamectin in an &lt;i&gt;embB&lt;/i&gt; deletion strain demonstrated that the target of selamectin is not cytosolic. Our data show that the concurrence of specific multiple mutations and complete integrity of the mycobacterial envelope are necessary for selamectin resistance. Our studies provide first-time insights into the antimycobacterial mode of action of the antiparasitic avermectins.IMPORTANCETuberculosis is the deadliest infectious disease of the 21&lt;sup&gt;st&lt;/sup&gt; century. New antibiotics are needed to improve treatment. However, developing new drugs is costly and lengthy. Drug repurposing is an alternative to the traditional drug discovery process. The avermectins are a family of drugs used to treat parasitic infections that are active against &lt;i&gt;Mycobacterium tuberculosis&lt;/i&gt;, the bacterium that causes tuberculosis. However, their mode of action in mycobacteria remains unknown. Understanding how avermectins kill mycobacteria can facilitate its development as an anti-mycobacterial drug, including against &lt;i&gt;M. tuberculosis&lt;/i&gt;.In this study, we used &lt;i&gt;Mycobacterium smegmatis&lt;/i&gt;, a non-pathogenic &lt;i&gt;M. tuberculosis&lt;/i&gt; surrogate model to understand the molecular mechanisms of how selamectin (a drug of the avermectin family selected for this study as a model) acts against mycobacteria. Our data show that the generation of resistance to selamectin is unlikely and that complete integrity of the mycobacterial envelope is necessary for selamectin resistance,","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0233224"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reliability of differential time to positivity technique for diagnosis of catheter-related bloodstream infections: a retrospective analysis.","authors":"Álvaro Irigoyen-von-Sierakowski, Marta Díaz-Navarro, Andrés Visedo, María Jesús Pérez-Granda, Pablo Martín-Rabadán, Patricia Muñoz, María Guembe","doi":"10.1128/spectrum.02678-24","DOIUrl":"https://doi.org/10.1128/spectrum.02678-24","url":null,"abstract":"<p><p>The differential time to positivity (DTTP) technique is the recommended conservative procedure to diagnose catheter-related bloodstream infection (C-RBSI). However, its reliability and accuracy remain under debate. Therefore, we aimed to compare the DTTP technique feasibility to detect C-RBSI compared to the catheter culture (CC) method. We conducted a 9-month retrospective study including bacteremic episodes in which both DTTP blood cultures (BC) and CC were obtained. We analyzed the diagnostic validity of the DTTP technique for detecting C-RBSI compared to the gold standard (C-RBSI with CC), along with patient clinical data. We included 37 episodes of C-RBSI where both DTTP BC and CC were obtained. C-RBSI was confirmed by both techniques in only 13 episodes (35.1%), whereas in 11 (29.7%) and 13 (35.1%), only DTTP BC or DTTP BC with CC (with a difference between catheter lumen and peripheral BC growth of <2 hours) was positive, respectively. Therefore, the validity values of the DTTP technique for predicting C-RBSI were as follows: sensitivity, 50.0%; specificity, 71.8%; positive predictive value, 54.2%; and negative predictive value, 68.3%. The distribution of microorganisms was similar among the three groups. All patients in whom colonization was not demonstrated by CC (<i>n</i> = 11) had been receiving antibiotics before catheter withdrawal. DTTP is a conservative technique that might help to diagnose C-RBSI mostly in situations where catheter removal cannot be achieved. However, it should be interpreted with caution and never be used to rule out C-RBSI. CC before starting antimicrobial therapy remains the most reliable method to diagnose and confirm an episode of C-RBSI.IMPORTANCEWe try to clarify the reliability of the differential time to positivity technique to predict C-RBSI. It may be interpreted with caution and considering clinical signs, as some C-RBSI can be misdiagnosed.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0267824"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The lack of specificity of <i>sodC</i>-based PCR for the detection of <i>Neisseria meningitidis</i> carriage in pharyngeal swabs from adolescents.","authors":"Dan Duy Pham-Tran, Lex E X Leong, Mark McMillan, Andrew Lawrence, Hassen Mohammed, Adam Finn, Helen S Marshall","doi":"10.1128/spectrum.02779-24","DOIUrl":"https://doi.org/10.1128/spectrum.02779-24","url":null,"abstract":"<p><p>The utility of the superoxide dismutase C (<i>sodC</i>) gene for detecting <i>Neisseria meningitidis</i> carriage in pharyngeal swab samples is unclear. While it has been explored as an alternative to the capsule transport A gene for detecting invasive strains, its potential as the sole target for real-time PCR in meningococcal carriage screening remains unexplored. This study aimed to assess the utility of using <i>sodC</i> as a target for detecting invasive and non-invasive strains of <i>Neisseria meningitidis</i> in pharyngeal swabs. Oropharyngeal swab samples collected from adolescents (approximately 15-18 years of age) enrolled in the \"B-Part-of-It\" cluster-randomized controlled trial (NCT03089086) were selected for this study. Samples were initially screened for the presence of specific <i>N meningitidis</i> porin A gene (<i>porA</i>) using PCR and then preserved by freezing at -80°C. An assay targeting the <i>sodC</i> gene was developed using the locked nucleic acid probe-based method. Upon optimization of the <i>sodC</i> assay, 1,092 samples were retested using this assay and compared against the results from the <i>porA</i> assays to determine concordance. Out of 1,092 samples tested, 965 (88.4%) were <i>sodC</i> positive, with 100% sensitivity and 13% specificity compared to <i>porA</i>. The positive predictive values and negative predictive values for the <i>sod</i>C assay were 12% and 100%, respectively. When compared to <i>porA</i> PCR as the gold standard, the <i>sodC</i> assay lacks sufficient specificity to serve as a stand-alone screening assay for the detection of <i>Neisseria meningitidis</i> carriage in pharyngeal samples from adolescents.IMPORTANCEWhile the <i>sodC</i> assay successfully detects <i>N. meningitidis</i>, we identified a limitation in its specificity due to potential cross-reactivity with other organisms, including <i>Haemophilus</i> spp., which can result in false positives. This limitation highlights the need for careful interpretation of <i>Neisseria meningitidis</i> carriage results, especially in epidemiological studies.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0277924"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spatial ecology of the <i>Neisseriaceae</i> family in the human oral cavity.","authors":"Jonathan J Giacomini, Julian Torres-Morales, Floyd E Dewhirst, Gary G Borisy, Jessica L Mark Welch","doi":"10.1128/spectrum.03275-24","DOIUrl":"https://doi.org/10.1128/spectrum.03275-24","url":null,"abstract":"<p><p>The human oral microbiome is a diverse ecosystem in which bacterial species have evolved to occupy specific niches within the oral cavity. The <i>Neisseriaceae</i> family, which includes human oral species in the genera <i>Neisseria</i>, <i>Eikenella</i>, <i>Kingella</i>, and <i>Simonsiella</i>, plays a significant role in both commensal and pathogenic relationships. In this study, we investigate the distribution and functional adaptations of <i>Neisseriaceae</i> species across oral habitats, focusing on their site tropisms and ecological roles. We employed a metapangenomic approach in which a curated set of reference genomes representing <i>Neisseriaceae</i> diversity was used for competitive mapping of metagenomic reads. Our analysis revealed distinct habitat preferences among <i>Neisseriaceae</i> species, with <i>Kingella oralis</i>, <i>Neisseria elongata</i>, and <i>Neisseria mucosa</i> primarily found in dental plaque; <i>Neisseria subflava</i> on the tongue dorsum; and <i>Neisseria cinerea</i> in the keratinized gingiva. Functional enrichment analyses identified genes and pathways underpinning habitat-specific adaptations. Plaque specialists showed metabolic versatility, with adaptations in nitrogen metabolism, including nitrate reduction and denitrification, lysine degradation, and galactose metabolism. Tongue dorsum specialists exhibited adaptations including enhanced capabilities for amino acid biosynthesis, short-chain fatty acid and glycerol transport, as well as lipopolysaccharide glycosylation, which may aid in resisting antimicrobial peptides and maintaining membrane integrity. These findings provide insights into the ecological roles and adaptive strategies of <i>Neisseriaceae</i> species within the human oral microbiome and establish a foundation for exploring functional specialization and microbial interactions in these niches.IMPORTANCEUnraveling the distribution and functional adaptations of <i>Neisseriaceae</i> within the human oral microbiome is essential for understanding the roles of these abundant and prevalent commensals in both health and disease. Through a metapangenomic approach, we uncovered distinct habitat preferences of various <i>Neisseriaceae</i> taxa across the oral cavity and identified key genetic traits that may drive their habitat specialization and role in host-microbe interactions. These insights enhance our understanding of the microbial dynamics that shape oral microbial ecology, offering potential pathways for advancing oral health research.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0327524"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel fluorescence immunoassay for the quantitative detection of HPV16 L1 antibodies in human serum samples using ZnCdSe/ZnS quantum dot-labeled antibodies.","authors":"Aiping Wang, Cheng Xin, Zhuting Chen, Jingming Zhou, Yumei Chen, Yankai Liu, Hongliang Liu, Chao Liang, Xifang Zhu, Yanhua Qi, Gaiping Zhang","doi":"10.1128/spectrum.01843-24","DOIUrl":"https://doi.org/10.1128/spectrum.01843-24","url":null,"abstract":"<p><p>Human papillomavirus type 16 (HPV16) is a high-risk virus linked to cervical cancer, primarily through its oncogenic proteins E6 and E7. The HPV16-L1 protein, the major capsid component, plays a key role in capsid formation and immune response. Monitoring anti-HPV16-L1 antibodies in serum is crucial for understanding infection dynamics and vaccine efficacy. This study aimed to develop a novel quantum dot-labeled blocking enzyme-linked immunosorbent assay (QDs-B-ELISA) for the quantitative detection of anti-HPV16-L1 antibodies. Monoclonal antibodies were produced and characterized against HPV16-L1 virus-like particles. A QDs-B-ELISA method was developed based on these antibodies and evaluated using 199 serum samples with previously established HPV16 status (\"known\" samples) and 170 serum samples with unknown HPV16 status at the time of testing (\"unknown\" samples). The diagnostic accuracy, sensitivity, specificity, and quantitative detection range of the QDs-B-ELISA were assessed and compared with commercial ELISA kits. The established QDs-B-ELISA exhibited high diagnostic accuracy (area under the curve, AUC = 0.9945), sensitivity (95.83%), and specificity (96.85%) for known serum samples. The lower limit of HPV16 antibody concentration detected by QDs-B-ELISA (0.0875 IU/mL) was considerably lower than that of the commercial ELISA kit, the Human Anti-HPV16-L1 Antibody (IgG) ELISA Kit (LS-F10262-1, Lsbio) (0.35 IU/mL), with a quantitative detection range of 13-1,737.8 IU/mL. When analyzing unknown human serum samples, the QDs-B-ELISA demonstrated a 97.06% agreement with commercial kits, and both inter-assay and intra-assay coefficients of variation were below 10%. The QDs-B-ELISA demonstrated high stability, sensitivity, and specificity, offering a valuable tool for surveillance and epidemiological studies of HPV16 infection.IMPORTANCEThis study introduces a novel quantum dot-labeled blocking enzyme-linked immunosorbent assay for detecting anti-HPV16-L1 antibodies, offering superior sensitivity and specificity compared to conventional methods. The improved performance enables more accurate HPV16 surveillance, epidemiological studies, and vaccine efficacy monitoring. This advancement may enhance early detection and risk assessment of HPV16 infections.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0184324"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anti-RBD immunoglobulin levels and their predictive value for SARS-CoV-2 neutralization.","authors":"Sabine Lichtenegger, Sissy Therese Sonnleitner, Sabine Saiger, Andrea Zauner, Melina Hardt, Barbara Kleinhappl, Gabriel E Wagner, Ivo Steinmetz","doi":"10.1128/spectrum.02148-24","DOIUrl":"https://doi.org/10.1128/spectrum.02148-24","url":null,"abstract":"&lt;p&gt;&lt;p&gt;A large number of studies have demonstrated that anti-receptor-binding domain (RBD)-binding antibody titers correlate with SARS-CoV-2 neutralization and protection from the disease. Unlike live virus neutralization assays, antibody-binding assays are easier to perform, require lower biosafety levels, and have therefore served as a substitute for virus neutralization assays throughout the SARS-CoV-2 pandemic. Although anti-RBD antibodies are usually neutralizing, there is evidence that they can also be non-neutralizing. Moreover, different immunization regimens can vary in the induction of SARS-CoV-2-neutralizing antibodies. In this study, we hypothesized that sera from individuals with different immunization and infection histories, but with the same amount of anti-RBD total immunoglobulin, differ in their neutralizing potency. A total of 27 sera from SARS-CoV-2 convalescent, vaccinated individuals, and 27 vaccinated-only individuals were investigated by using a widely used antibody-binding assay (Elecsys anti-SARS-CoV-2 S enzyme-linked immunosorbent assay [ELISA]) and a live virus neutralization assay. As expected, anti-RBD immunoglobulin units correlated with virus neutralization capacity within the vaccine and hybrid immunized group. However, sera from both groups with matched anti-RBD units varied significantly in their neutralization potential. In detail, our data indicate a significantly higher neutralization potency of hybrid immunity compared to vaccinated-only sera with similar anti-RBD immunoglobulin levels. Our study highlights the need for cautious interpretation of quantitative antibody data from anti-RBD ELISAs, especially when comparing differently immunized groups. In other words, very similar anti-RBD levels can show very different functional activity. This finding has implications for determining possible future correlates of protection.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Importance: &lt;/strong&gt;Throughout the SARS-CoV-2 pandemic, neutralizing antibody levels have been central to predict a protective immune response. Anti-receptor-binding domain (RBD) enzyme-linked immunosorbent assays (ELISAs) correlate with neutralization assays and are due to the integration of simple performance with timely results used as surrogate assays. However, previous studies determining correlation used homogeneous cohorts. We reevaluated the correlation of a frequently used anti-RBD ELISA and a live virus neutralization assay using a heterogeneous cohort consisting of a vaccinated group without prior SARS-CoV-2 infection and a vaccinated convalescent group. The neutralizing capacity of sera with matched anti-RBD units significantly differed between groups, decreasing the correlation of the assays. Our findings highlight the necessity of considering the immunization context when interpreting serological tests and suggest that different immunization groups may require distinct protective thresholds. Considering the immunization history, we can develop more accurate pre","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0214824"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803754","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Navigating the outbreak: a comprehensive analysis of pediatric <i>Mycoplasma pneumoniae</i> pneumonia via targeted next-generation sequencing in Wuhan, 2022-2023.","authors":"Changzhen Li, Jingjing Rao, Xiaomei Wang, Lifang Feng, Yun Xiang, Feng Tang","doi":"10.1128/spectrum.02463-24","DOIUrl":"https://doi.org/10.1128/spectrum.02463-24","url":null,"abstract":"<p><p>This study aims to delineate the epidemiological characteristics and impacts of the <i>Mycoplasma pneumoniae</i> (MP) outbreak, utilizing targeted next-generation sequencing (tNGS) to assess pathogen prevalence and co-infections in pediatric pneumonia cases. Pediatric patients admitted to Wuhan Children's Hospital with pneumonia from 1 October 2022 to 31 October 2023 were included. tNGS was used for comprehensive pathogen detection, including MP and other respiratory pathogens, with additional sequencing of the 23S rRNA gene V region to identify macrolide resistance mutations. This study enrolled 10,223 patients with pneumonia with a positivity rate of 98.4% by targeted next-generation sequencing. Fever (86.4%) and cough (79.3%) were the most common symptoms of <i>Mycoplasma pneumoniae</i> pneumonia (MPP). Lung consolidation (25.8%) was a common imaging feature, and corticosteroid use was noted in 22.5% of MPP patients. MP proved to be the primary pathogen, particularly evident during the MP pandemic, which began in March 2023 and peaked in October with a detection rate of 63.2%. Of the 4,397 MPP cases, 34.5% were sole infections, while 65.6% were co-infections, mostly with viruses. The main causative agents of co-infections were <i>Haemophilus influenzae</i> and <i>Rhinovirus</i>. The macrolide resistance rate was 79.03%. The A2063G mutation in the 23S rRNA V region is the dominant mutation. High-sensitivity C-reactive protein and serum amyloid A were significantly elevated in MPP, while the absolute counts of CD3⁺T, CD4⁺T, CD8⁺T, CD19⁺B, and NK cells were significantly reduced. This study demonstrates the utility of tNGS in identifying MP co-infections and macrolide resistance and highlights the role of inflammatory markers and lymphocyte subpopulations in differentiating MPP from non-<i>Mycoplasma pneumoniae</i> pneumonia for clinical management.IMPORTANCEOur study is of great scientific value as it provides practical solutions to clinical challenges and supports both clinical decision-making and public health policy. First, it presents new and important insights into the application of targeted next-generation sequencing (tNGS) technology, which enables rapid and accurate pathogen detection and overcomes the limitations of conventional diagnostic methods. Second, the large sample size, focusing specifically on children during a <i>Mycoplasma pneumoniae</i> epidemic, provides valuable epidemiologic data specifically for the Wuhan region. Finally, by integrating rapid tNGS detection with inflammatory markers and lymphocyte subsets, our study demonstrates direct clinical applications that have the potential to improve patient outcomes. This approach highlights the practical utility of our research in enhancing clinical decision-making and contributes important knowledge to the field.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0246324"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum for Gu et al., \"Dissemination of <i>bla</i>_NDM-harboring plasmids in carbapenem-resistant and hypervirulent <i>Klebsiella pneumoniae\"</i>.","authors":"Yihai Gu, Xiao Wang, Wei Zhang, Rui Weng, Qiucheng Shi, Xuan Hou, Hui Wang, Minghui Deng, Jian Mou, Yan Jiang","doi":"10.1128/spectrum.00577-25","DOIUrl":"https://doi.org/10.1128/spectrum.00577-25","url":null,"abstract":"","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0057725"},"PeriodicalIF":3.7,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum for Hernandez-Rodriguez et al., \"Strategies for genetic manipulation of the halotolerant black yeast <i>Hortaea werneckii</i>: ectopic DNA integration and marker-free CRISPR/Cas9 transformation\".","authors":"Yainitza Hernandez-Rodriguez, A Makenzie Bullard, Rebecca J Busch, Aidan Marshall, José M Vargas-Muñiz","doi":"10.1128/spectrum.00338-25","DOIUrl":"https://doi.org/10.1128/spectrum.00338-25","url":null,"abstract":"","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0033825"},"PeriodicalIF":3.7,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epitope mapping targeting the K205R protein of African swine fever virus using nanobody as a novel tool.","authors":"Xuedan Wei, Fengxia Zhang, Qiming Pei, Aijuan Shen, Duoxing Niu, Yaci Zhang, Ziheng Zhang, Yunshuo Lu, Angke Zhang, Gaiping Zhang, Hong Duan","doi":"10.1128/spectrum.01750-24","DOIUrl":"https://doi.org/10.1128/spectrum.01750-24","url":null,"abstract":"<p><p>African swine fever (ASF) is a highly infectious and lethal swine disease, leading to enormous losses in the pig industry. K205R, a non-structural protein of ASF virus (ASFV), is abundantly expressed at the early stages of viral infection and induces a strong immune response. In our previous study, five strains of K205R-specific nanobodies (Nbs) were screened through phage display technology, among which Nb1, Nb14, Nb35, and Nb82 exhibited good affinity. In the present study, the above four Nbs were successfully expressed in HEK293T cells and exhibited strong reactivity. Four Nbs recognized linear B-cell epitopes of K205R in both prokaryotic and eukaryotic expression systems. Besides, four Nbs specifically reacted with the K205R protein of ASFV-infected cells. Two epitopes ¹MVEPR⁵ and ¹⁸⁸RTQF¹⁹¹ were further identified, with highly conserved in different ASFV strains, and could interact with inactivated ASFV-positive sera, indicating that the two epitopes were natural linear B-cell epitopes. Moreover, structural analysis indicated that both epitopes were exposed on the surface of the K205R molecule. Notably, the identified epitope ¹⁸⁸RTQF¹⁹¹ was first reported. Overall, these findings provide valuable insights for K205R as an effective diagnostic tool and vaccine development.IMPORTANCEAfrican swine fever (ASF) is the number one killer affecting the pig industry, and there are no effective strategies for prevention. The ASFV K205R protein is prominently expressed in the early stages of viral infection, triggering a robust immune response. The full understanding of K205R protein epitopes provides a theoretical basis for the development of vaccine-candidate proteins. Nanobodies exhibit superior capability in detecting concealed epitopes of antigens compared with traditional antibodies. Here, we identify two epitopes ¹MVEPR⁵ and ¹⁸⁸RTQF¹⁹¹ based on nanobodies as a tool. Notably, the epitope¹⁸⁸RTQF¹⁹¹ is being reported for the first time. These epitopes are highly conserved in different ASFV strains and represent natural linear B-cell epitopes. This study opens up nanobodies as a new tool for the identification of epitopes and also provides a direct material basis for the development of ASFV vaccines.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0175024"},"PeriodicalIF":3.7,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143764298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}