Journal of Clinical Microbiology最新文献

{"title":"Advanced specificity and sensitivity studies relative to a research use only transcription-mediated amplification-based assay for <i>Treponema pallidum</i> RNA detection.","authors":"Trinity Krueger, Grace Kadonsky, Elizabeth Thelen, Amanda Zapp, Josephine Moore, Ahmet Muslu, Allan Pillay, Irene A Stafford, Erik Munson","doi":"10.1128/jcm.00388-25","DOIUrl":"10.1128/jcm.00388-25","url":null,"abstract":"<p><p>Preliminary experimentation has suggested that a research-use-only real-time transcription-mediated amplification assay for <i>Treponema pallidum</i> (RUO <i>T. pallidum</i> TMA) yields instances of <i>T. pallidum</i> nucleic acid detection that coincide with non-treponemal serology in men who have sex with men (MSM) at increased risk for sexually transmitted infection. To further characterize the specificity of RUO <i>T. pallidum</i> TMA testing, 3,586 rectal swab specimens reported as \"not detected\" by the assay generated a mean endpoint FAM fluorescence of 637.5 units. Introduction of <i>Treponema denticola</i>, <i>Treponema phagedenis</i>, and <i>Treponema refringens</i> nucleic acid into matrices generated mean endpoint FAM fluorescence ranging from 620.7 to 633.5 units (<i>P</i> ≥ 0.15 versus control). Introduction of lubricant to a pooled rectal swab matrix did not result in elevated FAM fluorescence (mean endpoint value 628.3 units [95% CI 612.0, 644.6]; <i>P</i> = 0.67). Introduction of talcum powder, urine, seminal fluid, and blood also failed to generate increased FAM fluorescence (<i>P</i> ≥ 0.20). Analytic sensitivity assessment was measured by serial 10-fold dilution of <i>T. pallidum</i> whole organism or <i>in vitro</i> 23S rRNA transcript in specimen transport medium or pooled rectal swab matrix and interrogation by the assay. Probit analysis estimated sensitivity (95% detection) of RUO <i>T. pallidum</i> TMA at 421-5,707 <i>in vitro</i> transcript copies/mL and 9-48 <i>T. pallidum</i> cells/mL, depending on dilution matrix. These results support RUO <i>T. pallidum</i> TMA as a highly sensitive method for <i>T. pallidum</i> detection that is not impacted by potentially cross-reactive organisms or interfering substances and may have adjunctive diagnosis capability for syphilis in MSM.</p><p><strong>Importance: </strong>Research-use-only <i>Treponema pallidum</i> transcription-mediated amplification (RUO <i>T. pallidum</i> TMA) has the potential to improve laboratory diagnosis of syphilis, particularly in patients at increased risk for sexually transmitted infection. The high analytic sensitivity and lack of cross-reactivity of the assay can facilitate other laboratories exploring the use of the test in a research setting.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0038825"},"PeriodicalIF":6.1,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144333165","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":"Development and evaluation of a Pan-Mucorales Real-time PCR and a multiplex Real-time PCR for detection and identification of <i>Rhizopus arrhizus, Rhizopus microsporus</i>, and <i>Mucor</i> spp. in clinical specimens.","authors":"Helen M Tang, Sharon C-A Chen, Kerri Basile, Catriona L Halliday","doi":"10.1128/jcm.01937-24","DOIUrl":"10.1128/jcm.01937-24","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Mucormycosis is a life-threatening infection associated with high morbidity and mortality. Rapid and accurate diagnosis is essential for improving patient outcomes. Conventional diagnostic methods, such as histopathology and culture, are limited by low sensitivity and prolonged turnaround times, while commercial polymerase chain reaction (PCR) assays are costly and may lack specific genus or species targets. Here, we present a novel molecular diagnostic workflow to facilitate the rapid detection of Mucorales directly from clinical specimens. This workflow integrates two in-house &lt;i&gt;in vitro&lt;/i&gt; diagnostic PCR assays: a real-time, qualitative Pan-Mucorales PCR, followed by a real-time multiplex genus/species-specific PCR targeting &lt;i&gt;Rhizopus arrhizus&lt;/i&gt;, &lt;i&gt;Rhizopus microsporus&lt;/i&gt;, and &lt;i&gt;Mucor&lt;/i&gt; spp. Specificity of the assays was validated using cultured isolates of Mucorales, as well as non-Mucorales fungi and bacteria. The diagnostic performance was assessed across 166 clinical specimens (70 Mucorales-positive and 96 negative), confirmed by an in-house panfungal PCR and DNA sequencing protocol. Specimens studied included fresh and formalin-fixed paraffin-embedded tissues, fluid, bronchoalveolar lavage/washing fluid, fine needle aspirate, cerebrospinal fluid, and bone. The Pan-Mucorales PCR demonstrated 98.6% sensitivity and 100% specificity, while the multiplex genus/species-specific PCR assay yielded sensitivities of 93.8% for &lt;i&gt;R. arrhizus&lt;/i&gt;, 70.8% for &lt;i&gt;R. microsporus&lt;/i&gt;, and 75% for &lt;i&gt;Mucor&lt;/i&gt; spp., each with 100% specificity. Concordance with the panfungal PCR (&gt;99% for Pan-Mucorales PCR and &gt;89% for multiplex PCR) was high, supporting the robustness of the workflow. This diagnostic approach has the potential to significantly reduce turnaround times, labor and costs, while streamlining the diagnostic process through timely, precise diagnostics.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Importance: &lt;/strong&gt;Mucorales fungi, identified collectively as a high-priority pathogen on the World Health Organization fungal priority pathogens list, are the causative agents of mucormycosis. Mortality is high (up to 80%), and early, accurate diagnosis is critical to enable timely initiation of targeted antifungal therapy and surgical debridement for source control to optimize patient outcomes. In our laboratory, as in many others, the current standard for the diagnosis of mucormycosis is histopathology and culture-based methods supplemented by panfungal PCR assay/DNA sequencing; however, this process may take 7 days, with considerable labor and cost implications. Here, we present two Mucorales-specific real-time PCR assays, which when used sequentially, reduce diagnostic turnaround time and costs to detect three common agents of mucormycosis-&lt;i&gt;Rhizopus microsporus&lt;/i&gt;, &lt;i&gt;Rhizopus arrhizus&lt;/i&gt;, and &lt;i&gt;Mucor&lt;/i&gt; species. This approach not only improves diagnostic efficiency and integration into workflow but can facilitate surveillance through accurate genus- and","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0193724"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153301/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144016790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preliminary performance of the VIDAS TB-IGRA as an aid in the diagnosis of individuals infected with <i>Mycobacterium tuberculosis</i>.","authors":"Potiandi Serge Diagbouga, Arthur Diakourga Djibougou, Camille Pease, Ariana Alcaide, Audrey Berthoux, Natalie Bruiners, Daniela Maria Cirillo, Ardjouma Combary, Nadine Falchero, Deborah Handler, Antoinette Kaboré, Alfred Lardizabal, Amanda Lopes, Marissa Loubet, Philippe Manivet, Clemence Margain, Valerie Meunier, Faiza Mougari, Alberta Onyuka, Sophie Rivoiron, Tani Sagna, Mathilde Sanvert, Léon Sawadogo, Jacques Simporé, Emmanuelle Cambau, Maria Laura Gennaro","doi":"10.1128/jcm.01641-24","DOIUrl":"10.1128/jcm.01641-24","url":null,"abstract":"<p><p>This preliminary study compares VIDAS TB-IGRA (bioMérieux, Marcy-l'Etoile, France) with the established QuantiFERON-TB Gold Plus (QFT-Plus) (Qiagen, Hilden, Germany) to evaluate diagnostic performance for the diagnosis of individuals infected with <i>Mycobacterium tuberculosis</i> complex (latent infection and disease). The study was multi-center and performed between 2 October 2019 and 4 February 2020. Participants were divided into tuberculosis (TB) disease, high-risk, and low-risk populations. The confirmed TB disease population included 104 patients. The high-risk population included 162 individuals with flagged risk factors on a questionnaire but without objective clinical confirmation of TB. The low-risk population included 117 healthy blood donors from the French National Blood Bank. Positive and negative percent agreement (PPA and NPA) were determined between the VIDAS TB-IGRA and QFT-Plus. In the TB disease population, sensitivity was measured against bacterial culture and PCR. The VIDAS TB-IGRA produced fewer indeterminate results than the QFT-Plus (1/104 vs 23/104) in the TB disease population and exhibited a sensitivity of 95.0% against bacterial culture. Furthermore, a 98.2% PPA was obtained in comparison to QFT-Plus. In the low-risk population, the VIDAS TB-IGRA demonstrated high specificity (94.9%) and a strong NPA (98.2%) compared to QFT-Plus. In the high-risk population, the VIDAS TB-IGRA exhibited a strong PPA (94.4%) with the QFT-Plus. A lower NPA was observed (85.2%) compared to QFT-Plus, which may be due to a higher sensitivity demonstrated in the TB disease population. The fully automated VIDAS TB-IGRA is a promising aid in the diagnosis of individuals infected with <i>Mycobacterium tuberculosis</i> (latent infection and active disease). It exhibits higher sensitivity while maintaining specificity and produces fewer indeterminate interpretations than QFT-Plus. Its easy-to-use, single-patient format may lead to increased TB testing to aid in the adequate diagnosis and management of the disease.IMPORTANCEThis study presents a comprehensive evaluation of the VIDAS TB-IGRA diagnostic test. This test is compared with the established QuantiFERON-TB Gold Plus to assess its effectiveness in diagnosing both latent and active tuberculosis (TB) infections. The study highlights the VIDAS TB-IGRA's higher sensitivity, fewer indeterminate results, and robust performance across different patient populations, including those with confirmed TB disease, high-risk, and low-risk groups. The findings suggest that the VIDAS TB-IGRA could enhance TB diagnosis and management, offering a fully automated, easy-to-use solution that reduces human error and result variability.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0164124"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153259/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144020881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of MALDI-TOF MS instruments and databases for the identification of uncommon yeasts, <i>Aspergillus</i> spp. and rare filamentous fungi.","authors":"Marion Dutkiewicz, Maéva Garros, Julie Bui, Véronique Charlier, Elodie Da Silva, Maryline Lemaire, Sarah Dellière, Anne-Cécile Normand, Renaud Piarroux, Samia Hamane, Théo Ghelfenstein-Ferreira, Alexandre Alanio","doi":"10.1128/jcm.01612-24","DOIUrl":"10.1128/jcm.01612-24","url":null,"abstract":"<p><p>Identification of uncommon fungi using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) remains challenging. Its performance depends on the protein extraction method, instrument, algorithm, and database. This study compared two established mass spectrometers (VITEK MS [bioMérieux] and Microflex [Bruker]) to the new VITEK MS PRIME (bioMérieux) and four databases, including MSI-2, a non-commercial database constructed with spectra acquired with Bruker instruments. Isolates of <i>Aspergillus</i> species, rare molds, and uncommon yeasts, previously identified by sequencing, were analyzed. After a two-step protein extraction, MALDI-ToF-MS identification was performed, and spectra were submitted to <i>in vitro</i> diagnostic (IVD) (KB3.2 and KB3.3 [bioMérieux]), research use only (RUO) (FilFungi V5 [Bruker]) databases and MSI-2. A total of 169 isolates (61 <i>Aspergillus</i> spp., 72 rare molds, and 36 uncommon yeasts), representing 33 genera and 96 species, were included. Identification rates at species level for all fungi were similar between VITEK MS and VITEKMS PRIME (79% vs. 77%). The main difference lies in the rates of non-analyzable spectra, being 15% (26 strains) for VITEK MS PRIME versus only 3% (5 strains) for VITEK MS. For <i>Aspergillus</i> and rare molds, MSI-2 performed equally well on VITEK MS and Microflex (92% vs. 91%), indicating a spectra compatibility. For uncommon yeasts, all databases performed equally at the species level. For <i>Aspergillus</i> and rare molds, FilFungi V5 performed poorly (23% and 21%), while MSI-2 was best (77% and 82%) due to broader species coverage. Misidentifications mostly involved cryptic <i>Aspergillus</i> species. MALDI-ToF-MS is a powerful tool for identifying rare fungi. Improvements are needed in completing commercial databases and optimizing acquisition systems for fungal spectra.</p><p><strong>Importance: </strong>Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) is key for fungal identification nowadays. We present here the largest comparison of MALDI-ToF-MS instruments and databases focusing on rare yeasts and molds identification, challenging the existing instruments and both commercial and academic databases. The strains were collected in Saint Louis Hospital (Paris) and identified using bar-code sequencing. We showed that commercial databases are efficient for the identification of the main fungal species tested, whereas the academic database outperforms them for the identification of cryptic species.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0161224"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153315/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Treatment and diagnostic challenges associated with the novel and rapidly emerging antifungal-resistant dermatophyte, <i>Trichophyton indotineae</i>.","authors":"Yasmeen Vincent Marbaniang, Daniela Leto, Huda Almohri, Mohammad Rubayet Hasan","doi":"10.1128/jcm.01407-24","DOIUrl":"10.1128/jcm.01407-24","url":null,"abstract":"<p><p><i>Trichophyton indotineae</i> is a recently discovered dermatophyte species that causes recalcitrant dermatophytosis. It was first reported from India and has quickly spread across the globe. The exact prevalence of <i>T. indotineae</i> remains unknown due to limited surveillance. It has reached epidemic proportions in the Indian subcontinent. In India, this new species has largely replaced other previously common dermatophytes. Reports from Western countries suggest most cases are imported, with some reports of local transmission. A recent report from the United Kingdom indicates that <i>T. indotineae</i> now accounts for 38% of dermatophyte isolates tested in their national referral laboratory. <i>T. indotineae</i> causes widespread, inflammatory dermatophytosis affecting large areas of the body. Dermatophytosis caused by <i>T. indotineae</i> is difficult to manage due to the limited availability of mycology laboratories capable of reliably identifying and performing antifungal susceptibility testing, and because of its resistance to commonly used antifungals. Culture and physiological characteristics cannot confirm identification to the species level, requiring species-level confirmation by molecular methods like internal transcribed spacer sequencing. It is important for clinicians and mycology laboratories to be aware of and consider the possibility of <i>T. indotineae</i> infection in patients with relevant demographic, clinical, and travel history. This would decrease delay in diagnosis, prevent inappropriate use of medications like steroids and ineffective antifungal agents, and provide opportunities to make recommendations for good hygiene practices to prevent transmission. In this mini review, we describe the emergence of <i>T. indotineae</i>, its diagnostic and treatment challenges, and the current state, and provide recommendations for future direction.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":"63 6","pages":"e0140724"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153305/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144266390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The dark matter of bacterial genomic surveillance-antimicrobial resistance plasmid transmissions in the hospital setting.","authors":"Annika Sobkowiak, Vera Schwierzeck, Vincent van Almsick, Natalie Scherff, Franziska Schuler, Kyrylo Bessonov, James Robertson, Dag Harmsen, Alexander Mellmann","doi":"10.1128/jcm.00121-25","DOIUrl":"10.1128/jcm.00121-25","url":null,"abstract":"<p><p>Dissemination of antimicrobial resistance (AMR) is a growing global public health burden. The aim of this study was to characterize AMR plasmid transmissions within a tertiary care hospital and identify relevant AMR plasmid transmission pathways. During an 18-month observation period, 540 clinical gram-negative multidrug-resistant bacterial (MDRB) isolates were collected during routine hospital surveillance and subjected to Pacific Biosciences long-read whole genome sequencing. Potential clonal transmissions were determined based on core genome multilocus sequence typing (cgMLST), and plasmid transmissions were detected using a novel real-time applicable tool for plasmid transmission detection. Potential transmissions were validated using epidemiological data. Among the 471 eligible MDRB isolates, we detected 1,539 plasmids; 84.41% of these were circularized. We identified 38 potential clonal transmissions in 24 clusters based on cgMLST and 121 potential plasmid transmissions in 24 clusters containing genetically related AMR plasmids. Among the latter clusters, 10 contained different multilocus sequence types (involving 2-38 isolates, median: 3 isolates), and nine contained multiple species (2-18 isolates, median: 4). Epidemiological data confirmed 19 clonal transmissions (in seven clusters) and an additional 12 plasmid transmissions (within eight plasmid clusters). Among these, we identified seven cases of intra-host and five patient-to-patient plasmid transmissions. We demonstrate that intra-host and patient-to-patient transmissions of AMR plasmids can be identified by combining long-read sequencing with real-time applicable tools during routine molecular surveillance. In addition, our study highlights that more than a decade of bacterial genomic surveillance missed at least one-third of all AMR transmission events due to plasmids.</p><p><strong>Importance: </strong>Antimicrobial resistance (AMR) poses a significant threat to human health. Most AMR determinants are encoded extra-chromosomally on plasmids. Although current infection control strategies primarily focus on clonal transmission of multidrug-resistant bacteria, until today, AMR plasmid transmission routes are neither understood nor analyzed in the hospital setting. In our study, we simultaneously determined both clonal, that is, based on chromosomes, and AMR plasmid transmissions during routine molecular surveillance by combining long-read sequencing with a novel real-time applicable software tool and validated all potential transmission events with epidemiological data. Our analysis determined not only the yet unknown plasmid transmissions within healthcare facilities or within the community but also resulted, in addition to the clonal transmissions, in at least a third more transmissions due to AMR plasmids.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0012125"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153326/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144002930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of MicroScan and VITEK 2 systems for susceptibility testing of Enterobacterales with updated breakpoints.","authors":"Sandra S Richter, Elizabeth L Dominguez, Aaron A Hupp, Matt Griffis, Shawn H MacVane","doi":"10.1128/jcm.00048-25","DOIUrl":"10.1128/jcm.00048-25","url":null,"abstract":"<p><p>We compared the performance of two commercial antimicrobial susceptibility testing (AST) systems for contemporary Enterobacterales strains using broth microdilution (BMD) as the reference standard with 2022 Clinical and Laboratory Standards Institute or current FDA breakpoints applied. Enterobacterales clinical isolates with BMD results were tested in parallel using VITEK 2 AST test cards (N802, XN15, bioMérieux, Hazelwood, MO, USA) and MicroScan NM56 AST panels (Beckman Coulter, Sacramento, CA, USA). The 200 isolates (57 <i>Escherichia coli</i>, 55 <i>Klebsiella pneumoniae</i>, 21 <i>Enterobacter cloacae</i> complex, 18 <i>Serratia marcescens</i>, 12 <i>Proteus mirabilis</i>, 10 <i>Citrobacter koseri</i>, 9 <i>Citrobacter freundii</i>, 8 <i>Klebsiella oxytoca</i>, 5 <i>Klebsiella aerogenes</i>, 3 <i>Providencia stuartii</i>, and 2 <i>Morganella morganii</i>) included 25% extended-spectrum beta-lactamase (ESBL), 23% carbapenem-resistant Enterobacterales (CRE), and 4.5% AmpC resistance phenotypes. For the 28 antimicrobial agents tested, essential agreement (EA, MIC within ±1 doubling dilution), categorical agreement (CA, same categorical interpretation: susceptible, intermediate, susceptible dose dependent, and resistant), and error rates were calculated using BMD as the reference standard. Time to results (TTR) was determined for each instrument for all 200 isolates. Hands-on time was assessed by timing two technologists each setting up six batches of five isolates on each system. Accuracy was similar between systems with an overall CA > 94% and EA ≥ 96%. The CA was ≥90% for most agents tested on both systems (exceptions were ampicillin-sulbactam, cefoxitin, minocycline, and nitrofurantoin). The MicroScan with Prompt inoculum preparation required less hands-on setup time than VITEK (1.29 vs 1.83 min/isolate). The median instrument TTR was less for VITEK (11.7 vs 18 hours, <i>P</i> < 0.001), yielding an overall faster turnaround time.IMPORTANCEThere are limited data directly comparing the performance of commercial antimicrobial susceptibility testing systems for contemporary bacterial strains using the Clinical and Laboratory Standards Institute broth microdilution method as the reference standard and applying updated breakpoints. These data will hopefully encourage labs to perform the necessary verification or validation studies needed to implement current breakpoints and ensure antimicrobial resistance is detected.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0004825"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153288/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063866","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photo Quiz: A cutaneous fungal infection with discordant biomarker results-a diagnostic challenge.","authors":"Guillaume Dumont, Sofiane Chaib, Emmanuelle Bontemps, Marjorie Brottier, Agnes Meybeck, Marjorie Cornu, Dea Garcia-Hermoso, Pierre Patoz","doi":"10.1128/jcm.02075-24","DOIUrl":"10.1128/jcm.02075-24","url":null,"abstract":"","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":"63 6","pages":"e0207524"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153258/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144266388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multicenter evaluation of the eQUANT system for use with disk diffusion AST of gram-negative bacteria directly from positive blood cultures.","authors":"Sanjucta Dutta, Michael R Jacobs, Caryn E Good, Ayman M Abdelhamed, Yun C Ying, Adrian Hoyos-Urias, Derrick J Sugui, Jacqueline U Anaeto, Eszter Deak, Rachael Valdez, Nitin K Rajan, Meike S Herget, Stefan Riedel","doi":"10.1128/jcm.01606-24","DOIUrl":"10.1128/jcm.01606-24","url":null,"abstract":"<p><p>Bloodstream infections frequently cause sepsis, a condition that can lead to organ dysfunction and death. Rapid antimicrobial susceptibility testing (AST) results are critical for appropriate medical intervention and to either de-escalate or escalate antibiotics to appropriate therapy. We describe the results from a multicenter clinical study evaluating the performance of the eQUANT system (Avails Medical, Inc., Menlo Park, CA). The eQUANT system generates a 0.5 McFarland equivalent suspension, the eMcFarland, directly from a positive blood culture bottle for use with downstream disk diffusion AST, thereby saving up to 24 hours compared to traditional AST workflow. A combination of fresh, prospectively collected clinical (42) and contrived (525) blood cultures were tested, and results for disk diffusion using the Avails eQUANT eMcFarland suspension as the inoculum were compared to disk diffusion results the next day using the standard 0.5 McFarland suspension prepared from plate subculture. Thirteen species of gram-negative rods were evaluated against 12 antibiotics. From the 2,679 pairs of AST results, overall categorical agreement was 95.0%. Overall very major errors, major errors, and minor errors were 0.15%, 0.60%, and 4.48%, respectively. The Avails eQUANT system has the potential to significantly accelerate the standard of care by eliminating the need for subculture.IMPORTANCERapid reporting of antimicrobial susceptibility test results for bacterial isolates from blood cultures is critical for timely implementation of optimal antimicrobial therapy and improves outcomes of sepsis patients. In this study, we demonstrate the accuracy and performance characteristics of the eQUANT system, which generates a 0.5 McFarland equivalent suspension, the eMcFarland, directly from a positive blood culture bottle for use with downstream disk diffusion antimicrobial susceptibility testing (AST). Using the eMcFarland suspension allows for accurate and standardized reporting of disk diffusion AST results from positive blood cultures.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0160624"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153300/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143995046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of disk diffusion susceptibility test methods for <i>Aerococcus</i> spp. and updates to Clinical and Laboratory Standards Institute MIC breakpoints.","authors":"Richard Maynard, Carmila Manuel, Synthia Simpkins, Michelle Haro, Nathan Ledeboer, Patricia J Simner, Mark Fisher, Romney Humphries","doi":"10.1128/jcm.00115-25","DOIUrl":"10.1128/jcm.00115-25","url":null,"abstract":"<p><p><i>Aerococcus</i> spp. are uropathogenic gram-positive bacteria that occasionally cause invasive infections. Minimal inhibitory concentration (MIC) breakpoints published by the Clinical and Laboratory Standards Institute (CLSI) were evaluated, and disk diffusion criteria developed, by testing 174 isolates of <i>Aerococcus</i> spp., by the CLSI M45 fourth edition workgroup. <i>Aerococcus urinae</i> growth was found to be significantly impacted by the brand of reference Mueller-Hinton broth base, and 14.6% of 117 <i>A</i>. <i>urinae</i> tested did not grow in cation-adjusted Mueller-Hinton broth supplemented with 5% lysed horse blood after 44 h incubation at 35°C in 5% CO₂ in any brand. Ampicillin and nitrofurantoin MIC breakpoints were established, and penicillin MIC breakpoints were adjusted, based largely on MIC distributions and pharmacokinetic data. Disk criteria were established for penicillin, ampicillin, vancomycin, nitrofurantoin, ciprofloxacin, levofloxacin, and tetracycline. Disk breakpoints could not be established for cefotaxime, ceftriaxone, or meropenem using standard CLSI disk masses due to large zones of growth inhibition that did not differentiate between MIC values and error rates that far exceeded CLSI standards. Trimethoprim-sulfamethoxazole breakpoints were also not possible to establish due to differences in thymidine content between MIC and disk diffusion testing media, which led to no correlation between MIC values and zones of growth inhibition for this antibiotic. The data summarized herein are the basis for proposed changes to the M45 guideline for <i>Aerococcus</i> spp., which will be published in the fourth edition.</p><p><strong>Importance: </strong><i>Aerococcus</i> species are an increasingly recognized cause of urinary tract infections and may cause serious infections, including endocarditis. Antimicrobial susceptibility testing (AST) is required for the management of these infections, as species within the genus display resistance to various antimicrobial agents. This study describes the data that informed updates to the Clinical and Laboratory Standards Institute M45 guideline for AST of fastidious or infrequently isolated bacteria. We describe the introduction of new ampicillin and nitrofurantoin breakpoints, updated MIC breakpoints for penicillin, and disk diffusion correlates for penicillin, ampicillin, ciprofloxacin, levofloxacin, nitrofurantoin, and tetracycline.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0011525"},"PeriodicalIF":6.1,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143993884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}