Journal of Clinical Microbiology最新文献

{"title":"Syphilis self-testing and implications for syphilis control and prevention.","authors":"Ayel Luis R Batac, Michael Marks, Joseph D Tucker, Rosanna Ŵ Peeling","doi":"10.1128/jcm.00982-25","DOIUrl":"https://doi.org/10.1128/jcm.00982-25","url":null,"abstract":"<p><p>The incidence of syphilis has risen dramatically in many regions despite the availability of affordable facility-based testing and curative treatment. The recent approval of over-the-counter syphilis self-tests (SSTs) represents an important advance for expanding diagnostic access and disease control and prevention. Evidence demonstrates that SSTs are accurate, usable, and acceptable. However, as with HIV and COVID-19 self-testing, implementation challenges remain, including ensuring equitable access, supporting vulnerable groups, securing linkage to care, and maintaining quality assurance.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0098225"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337028","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":"Self-collection for primary HPV screening using dry swabs: a review of clinical performance, laboratory considerations, and patient preferences.","authors":"Mondraya F Howard, Ria C Fyffe-Freil, Christina C Pierre, Henrietta O Maku, Stefania Di Costanzo, Thomas S Lorey, Dina N Greene","doi":"10.1128/jcm.00767-24","DOIUrl":"https://doi.org/10.1128/jcm.00767-24","url":null,"abstract":"<p><p>For cervical cancer screening, testing for high-risk human papillomavirus (hrHPV) detects more high-grade precancerous lesions, has a higher negative predictive value, and requires fewer lifetime screenings compared to cytology. As a result, both the US Preventive Services Task Force and the World Health Organization have endorsed hrHPV testing as the preferred screening method. Despite the utility, there are significant implementation challenges to adopting hrHPV primary screening across patients, providers, and institutions that must be addressed to ensure its widespread effectiveness. Here, we take a laboratory-centric approach to reviewing hrHPV primary screening, including discussion of specimen types and collection methods. For user experience, clinical and analytical validations, we focused on self-collected vaginal swab specimens stored dry during transport. Our analysis indicates that clinical laboratories should do their part to engage with institutional and clinical leadership to validate and promote the use of vaginal self-sampling for cervical cancer screening options within and outside the clinic. This work highlights the multiple studies that have validated dry swab collection as a simplified and high-quality method for hrHPV detection.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0076724"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336965","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":"Developing genome typing strategies for the emerging zoonotic pathogen <i>Streptococcus parasuis</i>.","authors":"Xiyan Zhang, Xueli Yi, Wenbo Luo, Jianping Wang, Chaoyuan Yuan, Wenfei Wei, Xuezhen Li, Jinhui Zhang, Han Zheng, Janguo Xu","doi":"10.1128/jcm.00741-25","DOIUrl":"https://doi.org/10.1128/jcm.00741-25","url":null,"abstract":"<p><p>The reported human infections with the emerging zoonotic pathogen <i>Streptococcus parasuis</i> are steadily rising. Rapid and standardized genotyping tools specific to <i>S. parasuis</i> are critically needed for epidemiological surveillance and identification of strains with zoonotic potential. This study developed a whole-genome sequence (WGS)-based typing strategy, encompassing average nucleotide identity, a minimum core genome (MCG) typing scheme, and a multilocus sequence typing (MLST) scheme using 255 <i>S</i>. <i>parasuis</i> genomes isolated from eight countries between the 1980s and 2024. The <i>S. parasuis</i> population was categorized into 12 MCG clusters based on 72,172 SNPs in non-recombining regions distributed across an MCG comprising 607 genes, forming two distinct lineages. The rapid MCG typing program accurately assigned 92.5% of <i>S. parasuis</i> genomes to their corresponding MCG clusters by identifying 4,509 cluster/subcluster-specific SNPs. To elucidate the clonal relationships among <i>S. parasuis</i> genomes, an MLST scheme was developed, defining 161 sequence types (STs) based on the allelic profiles of seven housekeeping <i>loci</i> (<i>aroA</i>, <i>cpn60</i>, <i>gki</i>, <i>mutS</i>, <i>sdhA</i>, <i>recA</i>, and <i>thrA</i>). Thirty-two STs that shared identical alleles at 6 <i>loci</i> were assigned to 10 complex clones, whereas 100 STs that shared identical alleles at 4 or more <i>loci</i> were grouped into 9 ST clades. The MCG typing scheme and the MLST scheme demonstrated sufficient discriminatory power, with Simpson's diversity index values of 0.8864 and 0.9821, respectively. This study characterized the <i>S. parasuis</i> population and provided a rapid, reproducible, and expandable WGS-based typing strategy for taxonomic identification, epidemiological surveillance, and evaluation of the zoonotic potential of <i>S. parasuis</i>.IMPORTANCEOur study provides valuable insights for developing effective prevention and control strategies for <i>Streptococcus parasuis</i> infections, by revealing the structural characteristics and phylogenetic relationship of <i>S. parasuis</i> population, by developing a whole-genome sequence-based typing strategy applicable for epidemiological surveillance, transmission investigation, and zoonotic potential evaluation.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0074125"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337014","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":"Validation of H5 influenza virus subtyping RT-qPCR assay and low prevalence of H5 detection in 2024-2025 influenza virus season.","authors":"David J Bacsik, Margaret G Mills, Luke D Monroe, Cassey Spring, Ailyn C Perez-Osorio, Jonathan C Reed, Ferric C Fang, Lori Bourassa, Pavitra Roychoudhury, Katharine H D Crawford, Kevin Snekvik, Alexander L Greninger","doi":"10.1128/jcm.00415-25","DOIUrl":"https://doi.org/10.1128/jcm.00415-25","url":null,"abstract":"<p><p>A sustained outbreak of H5N1 influenza virus among wild fowl and domestic livestock has caused more than 70 zoonotic infections in humans in North America, including two deaths. The United States Centers for Disease Control and Prevention has recommended rapid H5 subtyping for all hospitalized cases with influenza A virus infection to enable prompt initiation of antiviral treatment, as well as infection prevention and implementation of public health measures to control spread. To address these needs, we developed a qualitative multiplex RT-qPCR assay to subtype H5 influenza virus in nasal, nasopharyngeal, and conjunctival specimens with a limit of detection of 250 copies/mL. No cross-reactivity was observed with other common respiratory viruses, including seasonal H3N2 and H1N1 influenza A viruses. We retrospectively subtyped 590 influenza A virus-positive clinical specimens with Ct values less than 31 processed by University of Washington labs between March 2024 and February 2025, including 512 specimens collected during the 2024-2025 influenza season, and detected no H5 positives. After clinical implementation, we performed 150 clinically ordered H5 subtyping tests between February and April 2025 and again detected no positives. This work enhances clinical pandemic preparedness activities and highlights the exceedingly low prevalence of H5N1 influenza virus during the 2024-2025 respiratory season.IMPORTANCEThe spread of H5N1 influenza virus in the United States has led to the culling of almost 200 million birds, infected cow herds across 17 states, and resulted in 70 human infections as of July 2025. Rapid PCR subtyping of H5 influenza virus is critical to inform hospital infection prevention and public health to enable containment of viral transmission. Here, we report the design, validation, and clinical implementation of a qualitative multiplex H5-subtyping RT-qPCR assay for nasopharyngeal, nasal, and conjunctival swab specimens. Additionally, we offer the largest reported study of H5 subtyping of influenza A virus-positive specimens in the United States to date. No H5 infections were detected in 740 samples collected between March 2024 and April 2025 from patients with confirmed influenza A virus infection in a large academic medical system in Seattle, WA.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0041525"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336998","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":"Evaluation of a sample-to-answer real-time PCR assay for enterovirus detection in cerebrospinal fluid.","authors":"Grace Perkins, Jennifer Swink, Kevin Wade, Lori Hughes, Bijal A Parikh, Neil Anderson","doi":"10.1128/jcm.00864-25","DOIUrl":"https://doi.org/10.1128/jcm.00864-25","url":null,"abstract":"<p><p>Enteroviruses frequently cause aseptic meningitis, necessitating differentiation from bacterial meningitis to avoid unnecessary antimicrobial treatment and improve patient management. Polymerase chain reaction (PCR) remains the gold standard for enterovirus detection in cerebrospinal fluid (CSF) due to its high sensitivity and rapid turnaround time. The discontinuation of the Cepheid Xpert EV assay has created a gap in diagnostics. This study aimed to optimize two laboratory-developed real-time PCR assays using Diasorin direct amplification disc (DAD) and universal disc (UD) methods compared to the previously standard Cepheid Xpert EV assay. A total of 87 clinical CSF specimens were tested to assess sensitivity, specificity, and overall performance. There was no significant difference between the performance of the three assays compared to standard of care results. The Xpert EV and DAD methods had a 79.6% positive agreement, while the UD method had an 86.4% positive agreement. All methods achieved 100% negative agreement. Specificity testing against non-enteroviral pathogens confirmed no cross-reactivity. The estimated limit of detection was 100 copies/mL for Xpert EV, 250 copies/mL for UD, and 1,000 copies/mL for DAD, though clinical sensitivity remained high. Blood contamination affected the DAD assay but not the UD method due to its RNA extraction step. This study demonstrates the potential of DAD and UD methods as viable alternatives for rapid enterovirus detection in CSF, essential for patient management following the discontinuation of the Xpert EV assay. Further validation and comparison are needed for broader clinical applications.IMPORTANCERapid and accurate detection of enteroviruses in cerebrospinal fluid is crucial for patient management in cases of aseptic meningitis, especially following the discontinuation of the Cepheid Xpert EV assay. This study evaluates two laboratory-developed real-time PCR assays utilizing Diasorin direct amplification disc and universal disc methods, demonstrating their potential as viable alternatives for enterovirus detection with high sensitivity and specificity.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0086425"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337012","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":"Rapid detection of gram-negative antimicrobial resistance determinants directly from positive blood culture broths using a multiplex PCR system.","authors":"Stefanie Marxreiter, Jamie Marino, Katrina Callan, Judith Hargrave, Tricia Alston, Kathy Fauntleroy, Amy Robertson, Barry N Kreiswirth, Liang Chen, Mariana Castanheira, Matthew Hockin, Andrew C Hemmert, Amy Davis, Michael J Satlin, Lars F Westblade","doi":"10.1128/jcm.00384-25","DOIUrl":"https://doi.org/10.1128/jcm.00384-25","url":null,"abstract":"<p><p>Currently available rapid blood culture diagnostics detect few gram-negative resistance determinants, limiting their clinical utility. We prospectively evaluated the prototype BIOFIRE FILMARRAY Antimicrobial Resistance (AMR) Panel, a rapid multiplex PCR test that detects 31 AMR genes, on residual positive blood culture broths from patients with gram-negative bacteremia due to five target organisms at a New York City hospital. Predicted antimicrobial resistance based on the AMR Panel was compared to results from broth microdilution testing of bloodstream isolates recovered in culture. A simulated stewardship study assessed opportunities for the optimization of therapy if the AMR Panel results had been available for patient care in real time. We enrolled 148 patients with gram-negative bacteremia (<i>Escherichia coli, n</i> = 75; <i>Klebsiella pneumoniae, n</i> = 44; <i>Pseudomonas aeruginosa, n</i> = 17; <i>Enterobacter cloacae</i> complex<i>, n</i> = 9; and <i>Acinetobacter baumannii, n</i> = 3). The sensitivity of the AMR Panel for predicting antimicrobial resistance was ≥90% for 10/14 antimicrobial agents in <i>E. coli</i> and for 10/16 agents in <i>K. pneumoniae</i>. Specificity was ≥90% for 15/17 agents in <i>E. coli</i> and for all 16 agents in <i>K. pneumoniae</i>. Performance for other organisms was poor. For <i>E. coli</i> or <i>K. pneumoniae</i> bacteremia, use of the AMR Panel could have led to earlier escalation or de-escalation of β-lactam therapy in a majority of patients compared to what actually occurred. This study demonstrates that a rapid multiplex PCR test with a large menu of AMR genes can be applied to positive blood culture broths to rapidly predict resistance to frontline antimicrobial agents in patients with <i>E. coli</i> or <i>K. pneumoniae</i> bacteremia.IMPORTANCEPatients with gram-negative bacteremia require urgent treatment with antimicrobial agents that are effective against their infecting pathogen. However, conventional laboratory work-up of blood cultures takes days to yield results, and during this time, patients may receive ineffective therapies. We evaluated the prototype BIOFIRE FILMARRAY AMR Panel, an assay that detects 31 genes in gram-negative bacteria that confer resistance to β-lactams, fluoroquinolones, and aminoglycosides in approximately 1 hour, directly from positive blood culture broths, and compared these results to antimicrobial susceptibility testing of isolates recovered in culture. We found that the AMR Panel accurately predicted resistance in <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> to most antimicrobials. Moreover, if results from this assay had been used for patient care, there would have been opportunities to optimize antimicrobial prescribing more quickly than using conventional methods. These data demonstrate how novel molecular assays could optimize care for patients with <i>E. coli</i> and <i>K. pneumoniae</i> bacteremia.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0038425"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336977","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":"Detection of protozoan and helminth parasites in concentrated wet mounts of stool using a deep convolutional neural network.","authors":"Blaine A Mathison, Katie Knight, Jill Potts, Ben Black, John F Walker, Falon Markow, Amy Wood, Dustin Bess, Ken Dixon, Brian Cahoon, Weston Hymas, Marc Roger Couturier","doi":"10.1128/jcm.01062-25","DOIUrl":"https://doi.org/10.1128/jcm.01062-25","url":null,"abstract":"<p><p>Comprehensive diagnosis of gastrointestinal parasites is largely reliant on traditional stool microscopy, despite gains in molecular diagnostics. Wet-mount examinations remain a significant challenge for traditional microscopy, digital microscopy, and artifical intelligence (AI). We developed and validated a deep convolutional neural network (CNN) model that provides highly sensitive detection and presumptive classification of enteric parasites. Twenty-seven different parasites were trained on a CNN model using a wide diversity of 4,049 unique parasite-positive specimens (determined by traditional microscopy) collected in the USA, Europe, Africa, and Asia. Model validation was performed with a unique holdout set. In clinical validation, AI correctly detected 250/265 positive specimens (94.3% agreement) and 94/100 negative specimens (94.0%) before discrepant resolution. AI also detected 169 additional organisms from the validation specimens that were not previously identified. These additional detections underwent further discrepant analysis to adjudicate the results by scan review and microscopy. After resolution and inclusion of newly defined true positives and false positives, the positive agreement was 472/477 (98.6%). Negative agreement was variable by organism, ranging from 91.8% to 100%. A relative limit of detection study was performed comparing AI to three technologists of varying experience using serial dilutions of specimens containing <i>Entamoeba</i>, <i>Ascaris</i>, <i>Trichuris</i>, and hookworm. AI consistently detected more organisms and at lower dilutions of parasites than humans, regardless of the technologist's experience. The use of AI for wet-mount analysis is highly sensitive and detects considerably more organisms than traditional microscopy alone. The use of AI simplifies the parasitology workflow and reduces the reliance on traditional microscopy.IMPORTANCEGastrointestinal parasite ova and parasite (O&P) detection from stools is a manual, labor-intensive method requiring highly trained personnel. This testing has been largely unchanged in 100 years, with the exception of minor improvements in processing and fixation techniques. O&Ps are performed worldwide on millions of stool specimens a year, making any improvements in the process highly impactful. Digital slide imaging and artificial intelligence were recently established tools by our laboratory for improving permanent trichrome stain interpretation. This work builds on that breakthrough and describes the first comprehensive wet-mount AI model development and validation. Improved diagnostic yield, analytical sensitivity, and precision were demonstrated in this work through full clinical laboratory validation studies, including a specimen collection sourced from four continents and a diversity of fixatives and preparation techniques. This work represents the completion of a groundbreaking effort to bring parasite screening into the technological age.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0106225"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336985","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":"Dilution susceptibility testing method evaluation for the combination of ceftibuten and avibactam against Enterobacterales.","authors":"O N Walser, J Howland, K Jankowski, A Kennedy-Mendez, G G Stone, C M Pillar, D A Hufnagel","doi":"10.1128/jcm.00072-25","DOIUrl":"https://doi.org/10.1128/jcm.00072-25","url":null,"abstract":"&lt;p&gt;&lt;p&gt;With the increase in antimicrobial resistance and multidrug-resistant infections, the discovery of additional therapeutic options to combat different infection types and treat different patient populations is paramount. Many recently developed antibiotics are administered intravenously, which can needlessly overburden healthcare systems and may not be needed or appropriate for every patient. Extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales infections have increased since 2012 and are resistant to first-line therapies used for many infections including complicated urinary tract infections. To address the need for alternatives to intravenous therapies against ESBL-producing Enterobacterales, a novel combination of ceftibuten and a modified prodrug version of avibactam (ARX-1796) designed for oral administration is being developed to treat serious Gram-negative infections. Because clinicians often prescribe antibiotics to patients based on the susceptibility profile of the bacterium causing their infection-particularly when empiric therapy fails-establishing the parameters for the susceptibility testing is a vital phase of development. To address two key gaps in reference method development for CTB/AVI, this study follows CLSI M23 Tier 1 guidance to investigate the impact of nonstandard testing conditions on broth microdilution testing, as well as the correlation between broth microdilution and agar dilution methods. There was a high correlation between the methods for CTB/AVI (essential agreement ≥90%) when evaluating 153 Enterobacterales isolates, including recent clinical isolates with diverse antimicrobial resistance mechanisms. Besides 100-fold increased inoculum density and pH of 5.0, CTB/AVI activity was largely unaffected by 21 alternate broth microdilution test conditions.IMPORTANCENew antibiotics are desperately needed to combat expanding and new antimicrobial resistance trends globally. While there are antibiotics in the drug development pipeline that target antibiotic-resistant bacteria, most of these new antibiotics are not available in an oral formulation. Ceftibuten in combination with avibactam targets drug-resistant ram-negative organisms, including those that cause complicated urinary tract infections, and is orally administered. This combination fills a gap for clinicians seeking an appropriate oral therapeutic regimen for patients with drug-resistant infections. During anti-infective development, it is important to delineate the variables for susceptibility testing so that clinicians can confidently evaluate whether an infecting organism is resistant or susceptible to a potential therapy. This study evaluated dilution susceptibility testing methods for ceftibuten in combination with avibactam against targeted organisms (Enterobacterales) and found that broth and agar dilution testing methods agree, and this combination is recalcitrant to most variations, aside from low pH and inoculum size, in standard tes","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0007225"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336958","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":"Serodiagnosis of amoebic abscess: a retrospective diagnostic accuracy study of kits marketed in Europe.","authors":"E Prétot, M-P Brenier-Pinchart, P Tirard-Collet, F Gabriel, F Touafek, A Marteau, L Delcey, C Amiot, D Dupont, H Fricker-Hidalgo, H Sokol, A Moreno-Sabater, F Grenouillet","doi":"10.1128/jcm.00179-25","DOIUrl":"https://doi.org/10.1128/jcm.00179-25","url":null,"abstract":"<p><p>Amoebiasis is a cosmopolitan parasitic disease caused by <i>Entamoeba histolytica</i>. The most lethal form is extra-intestinal amoebiasis, mainly manifesting as liver abscesses. Diagnosis is based on clinical, radiological, and biological tests. However, there are currently few serology reagents still available on the European market for <i>in vitro</i> diagnosis, and comparative studies of existing reagents are required. We evaluated the performances of the four currently available reagents, either as standalone tests or in combination. Two enzyme-linked immunosorbent assays (ELISAs) for the detection of <i>E. histolytica</i> IgG were assessed: one manufactured by Bordier and the other by NovaTec. Additionally, an indirect hemagglutination technique, ELI.H.A <i>Amoeba</i>, and a latex particle agglutination technique, ELITex Bicolor <i>Amoeba</i>, both produced by ELITech Microbio, were evaluated. A total of 442 serum samples were selected from a shared centralized biobank of seven university hospitals in France. The samples included 79 from patients with amoebic abscess, 13 with amoebic colitis, and 350 from healthy donors and patients with parasitic and non-parasitic diseases, especially liver disease and immune dysfunction. The sensitivity of the four kits ranged from 87.3% to 97.5%, while their specificity ranged from 78.3% to 98.6%. Bordier ELISA demonstrated the highest sensitivity, while ELITex Bicolor <i>Amoeba</i> latex reagent exhibited the highest specificity. The combination of Bordier ELISA and/or ELI.H.A <i>Amoeba</i> for screening, combined with ELITex Bicolor <i>Amoeba</i> for confirmation of positive screening results, yielded the most optimal performance. This study highlights the benefits of large comparative studies of commercial assays to guide clinical microbiologists in their choices.</p><p><strong>Importance: </strong>Amoebiasis caused by <i>Entamoeba histolytica</i> is common in countries with low socio-economic levels. The most lethal form is extraintestinal amoebiasis, mainly liver abscesses, which require rapid and accurate diagnosis. Diagnosis is based on clinical, radiological, and, most importantly, serological tests. Implementing a diagnostic strategy requires both knowledge of how diagnostic tests compare with each other and thorough cross-validation. To help the clinical microbiologist choose a serological reagent, we evaluated the performance of four serological reagents currently available on the European market using a large biobank of sera from seven French university hospitals. In addition to patients with amoebic abscess and healthy donors, our study included many samples from patients with other parasitic and non-parasitic pathologies (liver diseases, immune dysfunctions) in order to study particularly non-specific reactivities.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0017925"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337008","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":"Performance evaluation of the QIAstat-Dx gastrointestinal panel 2 for the detection of <i>Clostridioides difficile</i> against multiple commercial assays.","authors":"Lavannya Sabharwal, Amorina Purpora, Derek Gerstbrein, Kennah Konrad, Brian Meisch, Nicolette Athanasiou, Ester Sala, Martí Juanola-Falgarona, Nestor Camenforte, Rebecca Smith-Aguasca, Pau Boher, Sarah Johnson, Matthew L Faron, Macy G Wood","doi":"10.1128/jcm.00576-25","DOIUrl":"https://doi.org/10.1128/jcm.00576-25","url":null,"abstract":"<p><p>Acute gastroenteritis is caused by a variety of pathogens, including bacteria, viruses, and parasites, which may have overlapping clinical signs and symptoms. Community-acquired gastroenteritis is caused by various pathogens, while healthcare-associated gastroenteritis is caused by <i>Clostridioides difficile</i>. Current diagnostic approaches utilize <i>C. difficile</i> nucleic acid amplification tests (NAATs) alone or in combination with an enzyme immunoassay for <i>C. difficile</i> antigen and toxin. This study evaluates the analytical performance of <i>C. difficile</i> target detection from 290 clinical specimens using the QIAstat-Dx multiplex gastrointestinal panel (GIP) 2 compared with four commercially available NAATs (Cepheid Xpert <i>C. difficile</i>, BD MAX Cdiff, Verigene CDF, and BioFire GI panel) as well as the <i>C. difficile</i> immunoassay. The positive percent agreement (PPA) and negative percent agreement (NPA) were calculated for all assays in the study. All molecular assays demonstrated a PPA greater than 85%, and an NPA greater than 96% when compared to the other NAATs. The QIAstat-Dx GIP 2 performance was comparable to the other NAATs, demonstrating 95.0%, 88.5%, 98.1%, and 93.7% PPA and 98.2%, 96.9%, 96.6%, and 99.1% NPA compared to Cepheid Xpert <i>C. difficile</i>, BD MAX Cdiff, Verigene CDF, and BioFire GI panel, respectively. Overall, the QIAstat-Dx GIP 2 assay was accurate and comparable to other commercially available methods, and the short assay setup time offers an efficient laboratory stool testing workflow.IMPORTANCE<i>Clostridioides difficile</i> infection ranges from diarrhea to severe, life-threatening pseudomembranous colitis. Clinical criteria include three or more liquid stools in 24 h, while diagnostic testing includes nucleic acid tests alone or in combination with an enzyme immunoassay for <i>C. difficile</i> antigen and toxin. This study evaluates the analytical performance of <i>C. difficile</i> detection from clinical specimens using the QIAstat-Dx multiplex gastrointestinal panel compared with four commercial assays (Cepheid Xpert <i>C. difficile</i>, BD MAX Cdiff, Verigene CDF assays, and BioFire GI panel) and a <i>C. difficile</i> immunoassay. The BioFire GI panel had the highest positive percent agreement (90-100%), followed by the QIAstat-Dx (88-98%), BD MAX (87-98%), and the Xpert assays (86-96%). The QIAstat-Dx demonstrated high positive and negative agreement with all nucleic acid tests. The short assay setup time for the QIAstat-Dx GIP 2 also offers a streamlined testing workflow.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0057625"},"PeriodicalIF":5.4,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145337010","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}