Journal of Clinical Microbiology最新文献

{"title":"An update on novel taxa and revised taxonomic status of bacteria isolated from human clinical specimens described in 2024.","authors":"Arianna Carella, Karen C Carroll, Erik Munson","doi":"10.1128/jcm.01068-25","DOIUrl":"https://doi.org/10.1128/jcm.01068-25","url":null,"abstract":"<p><p>Knowledge of novel and revised bacterial taxonomy facilitates routine operations in the medical microbiology laboratory and communication with important stakeholders. Notable new gram-positive taxa accepted in 2024 include the clinically significant <i>Staphylococcus brunensis</i> sp. nov., as well as <i>Streptococcus suis</i> subsp. <i>hashimotonensis</i> subsp. nov., which can be delineated from other <i>Streptococcus suis</i> subsp. by possession of the Lancefield group A antigen. Four novel <i>Enterobacterales</i> species are discussed, plus a novel eighth family <i>Gallaecimonadaceae</i>. One of these, <i>Providencia huashanensis</i> sp. nov. is clinically significant and harbors multiple genetic determinants that encode antimicrobial resistance mechanisms. <i>Stenotrophomonas forensis</i> sp. nov., derived from specimen transport medium, was determined to be a distinct component within the <i>Stenotrophomonas maltophilia</i> complex. <i>Bartonella tamiae</i> sp. nov. has become officially recognized as the agent of the first culture-confirmed bartonelloses in Thailand. Significant taxonomic revisions include the return of <i>Chlamydophila caviae</i> to the <i>Chlamydia</i> genus, creation of the novel <i>Metaclostridioides</i> genus for <i>Clostridioides mangenotii</i>, and reclassification of two <i>Kocuria</i> spp. into two subspecies of <i>Kocuria rosea</i>. Clinical updates are provided for 13 past <i>Journal of Clinical Microbiology</i> taxonomic compendium entries for which extensive clinical data were not previously available. These include potential clinical significance for non-<i>vaginalis</i> species of the <i>Gardnerella</i> genus and for <i>Pandoraea commovens</i> in a non-cystic fibrosis setting.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0106825"},"PeriodicalIF":5.4,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145308202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Valid and accepted novel bacterial taxa isolated from domestic companion and agricultural animals described in 2024.","authors":"Sara D Lawhon, Claire R Burbick, Trinity Krueger, Elena Ruiz-Reyes, Ayden Wisney-Leonard, Mallory Lenmark, Erik Munson","doi":"10.1128/jcm.01069-25","DOIUrl":"https://doi.org/10.1128/jcm.01069-25","url":null,"abstract":"<p><p>Recognizing and updating bacterial names is key to communication between the veterinary clinical microbiology laboratory, veterinarians, and clients. <i>Moraxella oculi</i> sp. nov., distinct from <i>Moraxella bovis</i> and <i>Moraxella bovoculi</i>, was isolated from a cow with infectious bovine keratoconjunctivitis. Several respiratory pathogens were recognized, including <i>Neisseria leonii</i> sp. nov. described from rabbits, <i>Mannheimia indoligenes</i> sp. nov. described from cattle in Europe, and <i>Moraxella haemolytica</i> sp. nov. described from a goat in China. <i>Stenotrophomonas forensis</i> sp. nov. is a novel designation within the <i>Stenotrophomonas maltophilia</i> complex associated with isolates derived from horses. New additions to the <i>Campylobacter</i> genus included <i>Campylobacter californiensis</i> sp. nov.<i>,</i> recovered from bovine feces during a raw milk-associated outbreak of campylobacteriosis in humans. Taxonomic revisions were most notable in Gram-positive organisms. A species within genus <i>Jeotgalicoccus</i> has been renamed, and the subspecies designation <i>Kocuria rosea</i> subsp. <i>polaris</i> comb. nov. has been created. <i>Streptococcus suis</i> was revised to <i>Streptococcus suis</i> subsp. <i>suis</i> subsp. nov. in recognition of the initial description of <i>Streptococcus suis</i> subsp. <i>hashimotonensis</i> subsp. nov., which was isolated from people in Japan with bite wounds from boars. <i>Chlamydophila caviae</i>, which causes respiratory disease and abortion in guinea pigs and is zoonotic, has been revised to <i>Chlamydia caviae</i> comb. nov.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0106925"},"PeriodicalIF":5.4,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145308204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Update on novel, validly published, and included bacterial taxa derived from nondomestic animals and taxonomic revisions published in 2024.","authors":"Claire R Burbick, Sara D Lawhon, Trinity Krueger, Elena Ruiz-Reyes, Mallory Lenmark, Ayden Wisney-Leonard, Erik Munson","doi":"10.1128/jcm.01070-25","DOIUrl":"https://doi.org/10.1128/jcm.01070-25","url":null,"abstract":"<p><p>Description of new and revised taxa originating from non-domestic animal species continues to be pursued. The majority are bacteria isolated from the alimentary system of a variety of healthy wildlife, adding to our understanding of microbial flora present in the normal state. A few new and revised taxa are associated with disease, including a novel <i>Erysipelothrix</i> species associated with sepsis in albatross and a revised species, <i>Allocoenonia anatina</i> comb. nov., associated with <i>Riemerella anatipestifera</i>-like disease in ducks and geese. Representative bacteria from genera that are additionally under study for the remediation of pollutants and bacteria that are related to significant animal or zoonotic pathogens are additionally discussed.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0107025"},"PeriodicalIF":5.4,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145308203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multicenter performance evaluation of the \"quanty TOXO (RH region)\" kit (Clonit) for molecular diagnosis of toxoplasmosis.","authors":"Céline Nourrisson, Emmanuelle Varlet, Juliette Guitard, Hélène Guegan, Cécile Nabet, Jean Menotti, Hervé Pelloux, Marie-Pierre Brenier-Pinchart, Yvon Sterkers","doi":"10.1128/jcm.00538-25","DOIUrl":"https://doi.org/10.1128/jcm.00538-25","url":null,"abstract":"<p><p>The diagnosis of congenital toxoplasmosis or disseminated toxoplasmosis in immunocompromised patients nowadays relies on molecular tools, in particular real-time PCR. There are many reagents on the market, and their evaluation by independent experts provides valuable information to medical biologists who are looking for a high-performance kit among the different references. Under the aegis of the French National Reference Center for Toxoplasmosis, we report here a multicenter evaluation of the analytical and clinical performances of the \"quanty TOXO (RH region)\" PCR assay manufactured by Clonit. The kit showed good analytical performance, as indicated by the results of serial dilution tests and external quality control samples. PCR efficiencies varied from 95% to 105%; linearity zone extended over four log units (<i>R</i>² >0.99), and limit of detection varied from <1 parasite/mL to between 1 and 5 parasites/mL, i.e., from <0.08 parasite/PCR to between 0.2 and 1 parasite/PCR, depending on the center. Based on 141 cryopreserved DNAs from a large range of clinical specimens, we determined a clinical sensitivity of 94.7% (71/75; 95% confidence interval [CI]: 87.1%-97.9%) and a clinical specificity of 100% (66/66; 95% CI: 94.5%-100%). Four false negative results were detected despite amplification carried out in duplicate. Overall, the \"quanty TOXO (RH region)\" PCR assay demonstrated satisfactory analytical and clinical performances for the diagnosis of toxoplasmosis, even using extraction and amplification techniques or biological matrices not validated by the manufacturer.IMPORTANCEDue to its speed and accuracy, PCR is now the gold standard for diagnosing congenital and disseminated toxoplasmosis. High-performance molecular testing is essential, especially for immunocompromised patients and congenital infections, to initiate early treatment. This diagnostic approach increasingly relies on commercial assays. However, commercially available kits do not guarantee performance. In this study, conducted by the French National Reference Center for Toxoplasmosis, we performed an independent multicenter evaluation of the \"quanty TOXO (RH region)\" PCR assay manufactured by Clonit. Our results showed that this kit delivered satisfactory results for routine diagnostic use. However, among the 141 clinical samples tested, four false negative results were noted, corresponding to specimens with low parasitic load.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0053825"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>In vitro</i> activity of antibiotic monotherapy and combination therapy with bacteriophages against <i>Staphylococcus aureus</i> LVAD-driveline infections.","authors":"Michèle M Molendijk, Nelianne J Verkaik, Corné P de Vogel, Nicole Lemmens-den Toom, Gwenan M Knight, Kadir Caliskan, Lonneke G M Bode, Annelies Verbon, Marion P G Koopmans, Miranda de Graaf, Willem J B van Wamel","doi":"10.1128/jcm.00272-25","DOIUrl":"https://doi.org/10.1128/jcm.00272-25","url":null,"abstract":"<p><p>Left-ventricular assist devices (LVADs) are increasingly used as a bridge to heart transplantation and destination therapy. These devices, especially the driveline, are susceptible to difficult-to-treat infections, associated with high morbidity and mortality rates. <i>Staphylococcus aureus</i> (<i>S. aureus</i>) is a major causative pathogen of LVAD infections. Antibiotic resistance and biofilm formation can complicate the treatment of these infections. A novel <i>in vitro</i> assay was developed to study the antibiotic susceptibility of <i>S. aureus</i> biofilm grown on LVAD drivelines. Besides antibiotic monotherapy, the effect of various antibiotics combined with rifampicin was studied. Additionally, we explored the efficacy of four individual phages and phage-antibiotic combinations as potential treatment strategies. Our data showed a decrease of susceptibility of the <i>S. aureus</i> biofilms to antibiotic monotherapy compared to planktonic <i>S. aureus</i>. With only rifampicin and erythromycin monotherapy resulting in full bacterial clearance. Combining antibiotics with rifampicin showed similar antimicrobial efficacy against <i>S. aureus</i> biofilms as rifampicin monotherapy. While both individual phages and a phage cocktail were effective against planktonic bacteria, phage efficacy was limited against <i>S. aureus</i> in biofilm. Combining phages with antibiotics did not clearly improve treatment efficacy, compared to antibiotic monotherapy. Contrarily, it even increased bacterial growth when phage administration preceded antibiotic treatment. Here, both antibiotic- and phage monotherapy showed reduced efficacy on LVAD-driveline biofilms. Additionally, phages did not show an additive value to antibiotic treatment of LVAD driveline infections. Further studies are needed to elucidate optimal treatment strategies for LVAD-driveline infections.IMPORTANCECurrent treatment strategies for <i>S. aureus</i> LVAD-driveline infections are based on <i>in vitro</i> antibiotic susceptibility of planktonic bacteria. However, LVAD infections are most often biofilm-related, which decreases antibiotic susceptibility significantly, resulting in discrepancies between <i>in vitro</i> antibiotic susceptibility and <i>in vivo</i> treatment success. Here, we have developed a novel <i>in vitro</i> assay to determine antibiotic susceptibility of <i>S. aureus</i> biofilm, grown in conditions relevant to LVAD-driveline infections. Next to antibiotic susceptibility, the susceptibility of this biofilm to bacteriophage mono- and combination treatment with antibiotics was evaluated as an alternative treatment strategy. In the future, this assay can be used to provide a better insight in <i>in vivo</i> antibiotic- and bacteriophage susceptibility of LVAD-driveline biofilms. Thereby improving <i>in vivo</i> treatment strategies for LVAD-driveline infections.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0027225"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive metabolomics combined with machine learning for the identification of SARS-CoV-2 and other viruses directly from upper respiratory samples.","authors":"Catherine A Hogan, Anthony T Le, Afraz Khan, LingHui David Su, ChunHong Huang, Malaya K Sahoo, Chieh-Wen Lo, Marwah Karim, Karin Ann Stein, Shirit Einav, Tina M Cowan, Benjamin A Pinsky","doi":"10.1128/jcm.02042-24","DOIUrl":"https://doi.org/10.1128/jcm.02042-24","url":null,"abstract":"<p><p>Metabolic profiling of respiratory samples from individuals infected and uninfected with respiratory viral infections may identify biomarker signatures that complement routine clinical diagnostic testing and offer unique insights into pathophysiology. We used liquid chromatography quadrupole time-of-flight mass spectrometry to generate untargeted metabolomic profiles and identified top biomarker signatures differentiating severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) positive from negative samples via machine learning. We then adapted these signatures to liquid chromatography-tandem mass spectrometry for targeted profiling and assessed classification performance, including samples positive for other respiratory viruses and negative for viral testing. A total of 1,226 samples were tested, including 521 positive samples for SARS-CoV-2, 97 for influenza A, 96 for respiratory syncytial virus (RSV), 211 for other respiratory viruses, and 301 negative samples. The top-performing model was the Light Gradient Boosting Model, which showed an area under the receiver operating characteristic curve (AUC) of 0.99 (95% confidence interval [CI], 0.99-1.00), sensitivity of 0.96 (95% CI, 0.91-0.99), and specificity of 0.95 (95% CI, 0.90-0.97). A separate machine learning analysis investigating the performance by viral subtype showed high performance for the identification of influenza A virus with an AUC of 0.97 (95% CI, 0.94-0.99) and RSV with an AUC of 0.99 (95% CI, 0.97-1.00). The two features with the highest ranking were identified as 3-oxo-heneicosanoic acid and 2-(4-hydroxyphenyl) ethanol. These findings extend our understanding of the metabolic impact of respiratory viral infections and support the potential of metabolomics to complement routine clinical diagnostic methods.IMPORTANCEMolecular testing has greatly improved how viruses are diagnosed; however, gaps remain, including limited sensitivity directly from specimens and inability to differentiate active from resolved infection. In this study, we investigated the use of a distinct diagnostic approach, mass spectrometry for detection of metabolites (small molecules) combined with machine learning analysis, for the diagnosis of SARS-CoV-2 and other respiratory viruses. We demonstrated strong performance of this approach directly from upper respiratory swab samples to differentiate SARS-CoV-2-infected versus uninfected individuals. Extension of this approach to influenza and RSV maintained a high level of performance. This research suggests that mass spectrometry-based infectious disease diagnostic testing has clinical potential and that these metabolomic features may reveal novel host-pathogen interactions and therapeutic targets. Applying a similar approach to prospective, multisite cohorts of patients with other infectious diseases carries potential to extend our understanding of the metabolic pathways involved in the host response to infection.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0204224"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251399","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Persistent multispecies dissemination of <i>armA</i>-carrying IncR plasmids among clinical and environmental bacterial populations in a Spanish veterinary hospital.","authors":"Carlos Serna, Mario Pulido-Vadillo, Bosco R Matamoros, Javier F Favieres, Natalia Montero, Claudia García Berdún, Marta E García, Jose L Blanco, Jose F Delgado-Blas, Bruno Gonzalez-Zorn","doi":"10.1128/jcm.00673-25","DOIUrl":"https://doi.org/10.1128/jcm.00673-25","url":null,"abstract":"<p><p>Aminoglycoside resistance mediated by 16S rRNA methyltransferases poses a growing threat in both human and veterinary medicine. Here, we conducted a retrospective genomic study of Enterobacterales isolates (<i>n</i> = 789) collected from clinical samples at a veterinary teaching hospital in Spain between 2011 and 2020. We identified four high-level aminoglycoside-resistant ST171 <i>Enterobacter hormaechei</i> subsp. <i>xiangfangensis</i> isolates carrying the <i>armA</i> gene, all from horses. Using Illumina and Nanopore sequencing, we determined that <i>armA</i> was located on a 70 kb IncR plasmid, also carrying other resistance genes such as <i>msr(E</i>), <i>mph(E</i>), <i>bla</i>_DHA-1, and <i>qnrB4</i>, embedded within a Tn<i>1548</i>-like element. Genomic comparisons indicated that the IncR plasmid was linked to a 2008-2010 ST11 <i>Klebsiella pneumoniae</i> outbreak in companion animals at the same hospital, with >95% sequence plasmid identity. Although the IncR plasmid lacked conjugative genes, it was mobilizable <i>in vitro</i> via co-resident conjugative plasmids. To probe for an environmental reservoir, we sampled three horse stalls in 2022 and recovered 19 <i>armA</i>-positive isolates (<i>E. hormaechei</i>, <i>Klebsiella pneumoniae,</i> and <i>Mixta calida</i>) whose IncR plasmids were nearly identical to that found in clinical clones. Broader analysis of 1,330 IncR plasmids from the genomic plasmid database PLSDB revealed that most were mobilizable, frequently co-integrated with other replicons, and carried diverse resistance genes, though <i>armA</i> was uncommon. These findings demonstrate that non-conjugative IncR plasmids can persist in the environment and be horizontally disseminated to clinical isolates over a long period of time, underscoring the need for routine, plasmid-focused genomic surveillance in veterinary healthcare settings within a One Health framework.IMPORTANCEThe spread of antimicrobial resistance threatens both human and animal health. In veterinary hospitals, bacteria can share resistance genes not only through direct transmission but also via mobile plasmids that persist in the environment. In this study, we uncovered a decade-long persistence of a non-conjugative IncR plasmid carrying the <i>armA</i> gene, which confers high-level aminoglycoside resistance, in a Spanish veterinary teaching hospital. This plasmid was found in clinical isolates of <i>Enterobacter hormaechei</i>, <i>Klebsiella pneumoniae,</i> and <i>Mixta calida</i> from horses and from the hospital environment. Our findings show that even plasmids lacking self-transfer capability can be maintained and disseminated across bacterial species over many years. These results highlight the need for routine genomic surveillance of plasmids in veterinary healthcare settings to prevent long-term environmental reservoirs from fueling recurrent outbreaks.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0067325"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251424","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plasma cell-free DNA PCR for diagnosing mucormycosis: are we there yet?","authors":"Valliappan Muthu, Inderpaul Singh Sehgal, Ritesh Agarwal","doi":"10.1128/jcm.01350-25","DOIUrl":"https://doi.org/10.1128/jcm.01350-25","url":null,"abstract":"","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0135025"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving time-to-result: head-to-head comparison of three rapid AST systems for Gram-negative bacteremia, including the newly developed VITEK REVEAL.","authors":"Damiano Squitieri, Giulia Menchinelli, Carlotta Magrì, Tiziana D'Inzeo, Barbara Fiori, Margherita Cacaci, Maurizio Sanguinetti, Giulia De Angelis, Brunella Posteraro","doi":"10.1128/jcm.01050-25","DOIUrl":"https://doi.org/10.1128/jcm.01050-25","url":null,"abstract":"<p><p>Early antimicrobial susceptibility testing (AST) in Gram-negative (GN) bloodstream infections is critical to guide appropriate therapy. This study evaluated three rapid AST (RAST) systems-VITEK REVEAL, direct-from-blood-culture VITEK 2 (VITEK 2-RAST), and EUCAST disk diffusion (DD-RAST)-using 220 prospectively collected GN-positive blood cultures (GN-PBCs). A total of 18 GN species were tested, including <i>Enterobacterales</i>, <i>Acinetobacter baumannii</i>, and <i>Pseudomonas aeruginosa</i>, against 25 antibiotics, four of which were next-generation β-lactam/β-lactamase inhibitor (BL/BLI) antibiotics. Reference broth microdilution was used for comparison. The VITEK REVEAL assay showed 97.1% essential agreement (EA), -7.7 bias, and 98.3% categorical agreement (CA) across 3,603 organism/antibiotic combinations. The VITEK 2-RAST assay showed 96.2% EA, -10.4 bias, and 98.4% CA across 3,941 combinations. The DD-RAST assay showed 98.2% CA across 2,388 combinations. Among 164 GN-PBCs tested with all three assays, EA rates were 97.5% for VITEK REVEAL and 96.3% for VITEK 2-RAST, with bias values of -7.1 and -6.9, respectively. Across all three assays, the CA rate was ≥98.2%, and very major error (VME) rates were ≤1.8%. Mean time-to-result (TTR) was significantly shorter with VITEK REVEAL (6 h 32 min) compared to VITEK 2-RAST (13 h 51 min) and the fixed 8 h of DD-RAST. In resistant organisms, VITEK REVEAL produced significantly faster results than in susceptible or intermediate ones, including BL/BLI antibiotics. <i>A. baumannii</i> yielded the shortest TTR, with no observed VME. These results support the use of RAST directly on GN-PBCs, with VITEK REVEAL emerging as a particularly promising option for rapid and accurate testing.</p><p><strong>Importance: </strong>This study is the first to conduct a direct comparison of three rapid antimicrobial susceptibility testing (RAST) systems-VITEK REVEAL, VITEK 2-RAST, and DD-RAST-on a large, prospectively collected cohort of Gram-negative-positive blood cultures (GN-PBCs). The data offer valuable insights for laboratories evaluating RAST implementation, especially in EUCAST-based contexts. Among the systems tested, VITEK REVEAL stood out for its combination of rapid turnaround and high accuracy, including for antibiotic-resistant organisms and β-lactam/β-lactamase inhibitor (BL/BLI) antibiotics. These findings underscore the potential of RAST systems to deliver timely and reliable susceptibility results directly from GN-PBCs, thereby supporting more effective antimicrobial stewardship and clinical decision-making. However, the real-world impact will also depend on how well these systems integrate into routine workflows, as seamless implementation can directly affect time-to-result (TTR)-an essential element in optimizing the management of GN-bloodstream infection (BSI).</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0105025"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An in-house <i>Kingella kingae</i> PCR in a large pediatric cohort: impact on diagnosis, antimicrobial stewardship, and clinical outcomes.","authors":"Sophonie Jean Oyeniran, Amy L Leber, Huanyu Wang","doi":"10.1128/jcm.00986-25","DOIUrl":"https://doi.org/10.1128/jcm.00986-25","url":null,"abstract":"<p><p><i>Kingella kingae</i> is a bacterial pathogen associated with bone and joint infections and a major pathogen in pediatric populations, particularly in those ≤5 years of age. Molecular detection methods have demonstrated significant increases in sensitivity compared to culture, allowing rapid and accurate identification. The objective of this study was to characterize the performance of an in-house <i>K. kingae</i> PCR (KKIN PCR) and determine its impact on antimicrobial utilization and clinical management in a pediatric population. Laboratory records from 1 September 2014 to 31 January 2024 were reviewed to identify subjects ≤18 years old who had KKIN PCR testing performed at Nationwide Children's Hospital. Demographics, symptoms, radiologic and laboratory findings, hospitalization status, antimicrobial therapy, and clinical outcomes were analyzed. During the study period, 500 unique subjects with KKIN PCR ordered had complete data available, among whom 46 had <i>K. kingae</i> detected by PCR or culture. Compared to the clinical diagnosis, KKIN PCR had sensitivity, specificity, and positive and negative predictive values of 95.7, 100, 100, and 99.6%, respectively. Thirty-six (80%) subjects were detected by PCR alone, improving case identification by fourfold compared to culture (<i>n</i> = 9). Among subjects ≤5 years old, <i>K. kingae</i> was the most common organism detected. The median time to result for KKIN PCR was 26.5 h, and 38 (84%) subjects had antimicrobial modifications within 1 day of KKIN PCR report, resulting in de-escalation or antimicrobial optimization. This study demonstrates that routine use of in-house KKIN PCR significantly increased the diagnosis, facilitated timely and targeted treatment, and improved management of pediatric bone and joint infections.IMPORTANCE<i>Kingella kingae</i> (KKIN) has long been recognized as a major cause of bone and joint infections in pre-school aged children. However, diagnosis and prevalence of KKIN infection are underestimated due to poor culture recovery. Previous studies have shown that molecular-based methods improve KKIN detection compared to culture, but these methods are not widely implemented or routinely used in clinical microbiology laboratories. This study describes the performance and clinical utility of an in-house laboratory-developed KKIN PCR in a large pediatric cohort over a nearly 10-year period. In addition to demonstrating improved KKIN detection compared to culture, it also shows that rapid availability of in-house KKIN PCR facilitated timely antimicrobial de-escalation and potentially contributed to shortened hospital length of stay compared to previous reports. This study highlights a critical diagnostic gap that can be alleviated with a validated laboratory-developed PCR to improve diagnosis and management of KKIN infections.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0098625"},"PeriodicalIF":5.4,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}