Journal of Clinical Microbiology最新文献

{"title":"Update on North American tick-borne diseases and how to diagnose them.","authors":"Kyle G Rodino, Elitza S Theel, Bobbi S Pritt","doi":"10.1128/jcm.00807-23","DOIUrl":"10.1128/jcm.00807-23","url":null,"abstract":"<p><p>Recent decades have seen a rise in the incidence of tick-borne diseases in the US, along with an increased number of pathogens transmitted by ticks, and geographic expansion of tick populations. A variety of laboratory testing methodologies are available for the diagnosis of tick-borne diseases, including serology, microscopy, and molecular-based methods. The preferred approach varies by the specific disease, locally available test options, and the stage of illness at patient presentation. This mini-review focuses on updates in our understanding of the epidemiology of tick-borne diseases in the US and advances in the field of laboratory diagnostics.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0080723"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345162/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144608541","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":"Enhancing diagnostic preparedness for H5N1: a validation study of H5 single-plex assay and detection across multiple platforms.","authors":"Yuan Chao Xue, Jennifer Bertsch, Kaylin Monacy, Carter Haynes, Natalie Williams-Bouyer, Barbara M Judy, Patrick C Newman, Thomas G Ksiazek, Lyudmyla V Marushchak, Gregory C Gray, Ping Ren","doi":"10.1128/jcm.00681-25","DOIUrl":"10.1128/jcm.00681-25","url":null,"abstract":"<p><p>The increasing transmission of highly pathogenic avian influenza (HPAI) H5N1 from animals to humans underscores the urgent need for enhanced diagnostic capabilities in clinical microbiology laboratories. Although <i>in silico</i> analysis suggests that commercial multiplex respiratory panels can detect H5N1, these assays lack subtyping H5N1 capabilities, and their real-world performance remains largely unverified. In this study, we evaluated the limit of detection (LoD) for H5N1 using three commercial molecular diagnostic assays routinely employed at our institution: bioMérieux BioFire Respiratory 2.1 Panel, Cepheid Xpert Xpress CoV-2/Flu/RSV Plus, and Hologic Panther Fusion SARS-CoV-2/Flu A/B/RSV assays. All three reliably detected H5N1 were at low viral concentrations. To enable H5N1 detection, we also developed a real-time RT-PCR H5 single-plex assay on the Hologic Panther Fusion Open Access platform for reflex testing of influenza A-positive specimens. In conclusion, although current commercial assays lack influenza A H5 subtype differentiation, our validation data provide critical performance information. When integrated with a targeted H5 assay, these tools can enhance clinical decision-making and public health surveillance by reducing the risk of missed H5N1 infection cases.IMPORTANCEThis study addresses a growing public health concern: the spread of bird flu (H5N1) from animals to humans. Most hospital laboratories use commercial tests to detect respiratory viruses like the flu, but these tests cannot tell if someone has the specific and more dangerous H5N1 strain. To help solve this, we tested three commonly used diagnostic tools and found that they can detect H5N1 even at low levels. However, since they cannot identify the specific H5 subtype, we also developed and validated a follow-up test that runs on one of the existing laboratory high-throughput equipment. This test can confirm whether a patient infected with the flu has the H5N1 strain. By combining these tools, hospital laboratories can improve early detection of H5N1, support better patient care, and help public health officials respond more effectively to outbreaks.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0068125"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345207/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144659366","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 Brief Case: <i>Legionella micdadei</i> caught red-handed.","authors":"Priyanka Gupta, Hiranmayi Vemaganti, Nahed Ismail, Vijayalakshmi Viju Ananthanarayanan","doi":"10.1128/jcm.00302-25","DOIUrl":"10.1128/jcm.00302-25","url":null,"abstract":"","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":"63 8","pages":"e0030225"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345241/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144835256","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":"Isolation and characterization of avian metapneumovirus subtypes A and B associated with the 2024 disease outbreaks among poultry in the USA.","authors":"Jianqiang Zhang, Liying Tian, Jaime Dittman, Baoqing Guo, Kay Kimpston-Burkgren, Erin Kalkwarf, Eman Gadu, Phillip Gauger, Mohamed El-Gazzar, Yuko Sato","doi":"10.1128/jcm.00333-25","DOIUrl":"10.1128/jcm.00333-25","url":null,"abstract":"<p><p>Avian metapneumovirus (aMPV) is currently classified into four subtypes (aMPV-A, -B, -C, and -D). In late 2023 and early 2024, aMPV-A and aMPV-B were detected in US poultry for the first time, causing significant economic losses. This study analyzed aMPV RT-PCR data from 2,204 samples (1,158 turkey, 936 chicken, and 110 other breeds) submitted to a US veterinary diagnostic laboratory between January and November 2024. A higher percentage of turkey samples (51.04%) tested PCR-positive for aMPV-A and/or aMPV-B compared to chicken samples (15.6%), with aMPV-A showing an overall higher positive rate than aMPV-B, although the positive rates varied by state. Additionally, four aMPV-A and three aMPV-B isolates were successfully recovered from clinical samples using primary chicken embryo lung and/or fibroblast cells. Two aMPV-A isolates (USA/IA55601-6/2024 and USA/IA56509-5/2024) and two aMPV-B isolates (USA/NC20487-GA/2024 and USA/NC23734-GA/2024) were adapted to grow efficiently in a Vero cell line, reaching titers of ~10⁴-10⁶ TCID₅₀/mL between passages 4 and 10 for aMPV-A and between passages 1 and 10 for aMPV-B. Whole genome sequencing of the two aMPV-A and two aMPV-B isolates at different passages revealed that the viruses progressively acquired several nucleotide changes, some of which led to amino acid substitutions in different viral proteins during 10 passages in cell culture. Comparative analysis with 46 aMPV-A, -B, -C, and -D GenBank sequences showed that US aMPV-A and aMPV-B strains were genetically closely related within their subtypes. These cell culture-adapted US aMPV-A and aMPV-B isolates provide valuable tools for further characterization of aMPV and vaccine development.</p><p><strong>Importance: </strong>Avian metapneumovirus (aMPV) subtypes A and B were first detected in US poultry in late 2023 and early 2024, rapidly spreading nationwide and posing a significant threat to the industry. This study analyzed RT-PCR data from 2,204 clinical samples (January to November 2024) to determine aMPV-A and aMPV-B detection rates across poultry species, age groups, and states, providing insights into their epidemiology in the USA. Modified live vaccines are urgently needed to control aMPV but are hindered by the lack of US isolates growing efficiently in cell culture. We successfully isolated aMPV-A and aMPV-B in primary chicken embryo cells and adapted them to a Vero cell line. Their infectious titers and genetic stability were characterized over serial passages. These US aMPV-A and aMPV-B cell culture isolates provide valuable tools for studying pathogenesis, determining virus infectious doses, evaluating disinfectants and antivirals, and developing vaccines.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0033325"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345154/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144600612","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":"Validation and multi-site deployment of a lyophilized qRT-PCR reagent for the molecular diagnosis of avian influenza and rabies in Sub-Saharan African regions.","authors":"Petra Drzewnioková, Irene Brian, Marzia Mancin, Andrea Fortin, Morgane Gourlaouen, Angélique Angot, Mamadou Niang, Isaac Dah, Kouramoudou Berete, Adama Diakite, Fatou Tall Lo, Clement Meseko, Emilie Go-Maro, Valeria D'Amico, Viviana Valastro, Baba Soumare, Paola De Benedictis, Isabella Monne, Valentina Panzarin","doi":"10.1128/jcm.00080-25","DOIUrl":"10.1128/jcm.00080-25","url":null,"abstract":"<p><p>Molecular methods are widely accepted as gold-standard techniques for the laboratory diagnosis of most human and animal pathogens. However, most molecular protocols rely on reagents that need to be transported and stored at a freezing temperature, a requirement that might affect their reliability in areas where the cold chain cannot be guaranteed. Over the years, several lyophilized molecular products have been marketed to circumvent this issue. We therefore evaluated the feasibility of replacing liquid reagents with freeze-dried formulations for the molecular diagnosis of avian influenza (AIV) and rabies (RABV) viruses, two priority zoonotic pathogens widely spread in Sub-Saharan Africa. Among six different commercial freeze-dried kits, we selected one reagent due to its easy-to-use features, single-reaction format, and preliminary performance assessment. Through a more in-depth evaluation, we determined its analytical and diagnostic performance and formulation stability and obtained results comparable to those of standard liquid master mixes. However, for the detection of divergent lyssaviruses, the lyophilized reagent's sensitivity was affected by suboptimal complementarity between the oligonucleotides and target sequences. Finally, a multi-site evaluation in four veterinary diagnostic laboratories located in Sub-Saharan Africa demonstrated the successful deployment of AIV and RABV assays utilizing the freeze-dried reagent, which can interchangeably replace liquid qRT-PCR/RT-PCR kits. Altogether, our results indicate that the investigated lyophilized master mix represents a valid alternative to liquid reagents for the molecular diagnosis of avian influenza and rabies and has the potential for broader applications to other relevant infectious diseases upon proper validation.IMPORTANCEMolecular diagnostic protocols rely on reagents that need to be transported and stored at freezing temperatures. Meeting this requirement can be challenging in areas where the maintenance of the cold chain is not guaranteed, such as in Sub-Saharan Africa. Our study aimed to assess the feasibility of using lyophilized reagents as a replacement for liquid reagents in the molecular diagnosis of two widespread zoonotic pathogens in Sub-Saharan Africa, namely, avian influenza and rabies. To accomplish this, we selected a commercially available lyophilized reagent based on its format and performance characteristics. We conducted a laboratory validation to assess the use of the lyophilized reagent throughout the entire diagnostic process. We also conducted a reproducibility test involving African laboratories as potential end-users. Our findings confirm that the lyophilized reagent can replace traditional liquid reagents to diagnose rabies and avian influenza and suggest its possible use for a wider range of infectious diseases after undergoing appropriate validation.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0008025"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345159/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144540377","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":"Performance evaluation of early growth isolates for automated and manual broth microdilution antimicrobial-susceptibility testing.","authors":"Lucas J Osborn, Lindsay Osborn, Irvin Ibarra-Flores, Marisol Garcia, Kaitlyn Perez, Ali Farhadiayoubloo, Melissa Mitrou, Cristina Costales, Jennifer Dien Bard","doi":"10.1128/jcm.00236-25","DOIUrl":"10.1128/jcm.00236-25","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Prolonged turnaround times (TAT) represent a major limitation to current automated susceptibility testing systems and manual susceptibility-testing methods such as broth microdilution. As a result, targeted therapy for patients may be delayed, portending suboptimal clinical outcomes. One contributing factor is the 18-24 h of incubation prior to antimicrobial-susceptibility testing (AST) recommended by the Clinical Laboratory Standards Institute (CLSI) and some automated AST manufacturers. This study evaluates the performance of AST by manual broth microdilution (Thermo Fisher Sensititre) and an automated AST system (BD Phoenix) on isolates incubated for 6 h (early growth AST, egAST) compared with 18-24 h (standard growth AST, sgAST). An initial proof-of-concept study conducted on gram-negative (&lt;i&gt;n&lt;/i&gt; = 5) and gram-positive (&lt;i&gt;n&lt;/i&gt; = 2) quality control strains incubated for 6 or 24 h prior to Sensititre and Phoenix demonstrated 100% essential agreement. Subsequently, we evaluated the performance of egAST on gram-positive (&lt;i&gt;n&lt;/i&gt; = 49, Phoenix; &lt;i&gt;n&lt;/i&gt; = 46 Sensititre) and gram-negative (&lt;i&gt;n&lt;/i&gt; = 81 Phoenix; &lt;i&gt;n&lt;/i&gt; = 61 Sensititre) patient-derived isolates with diverse resistance profiles compared with sgAST. In total, 1,666 organism-drug combinations were tested by Sensititre (560 gram-positive and 1,106 gram-negative) and 1,927 by Phoenix (409 gram-positive and 1,518 gram-negative). For &lt;i&gt;Enterobacterales&lt;/i&gt;, egAST using Phoenix revealed 1.3% minor errors (MiE), 0.17% major errors (ME), and 1.1% very major errors (VME) compared with sgAST. Phoenix egAST performance for &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; revealed 0.18% MiE and no ME or VME. Similarly, egAST of &lt;i&gt;Enterobacterales&lt;/i&gt; by Sensititre revealed 1.5% MiE and no ME or VME, whereas 2.8% MiE, 6.6% ME, and no VME were observed for &lt;i&gt;P. aeruginosa&lt;/i&gt;. For &lt;i&gt;Staphylococcus&lt;/i&gt; spp. and &lt;i&gt;Enterococcus&lt;/i&gt; spp., there were no MiE, ME, or VME on the Phoenix system, whereas early growth Sensititre showed 3.14% MiE, 0.3% ME, and 2.99% VME for &lt;i&gt;Staphylococcus&lt;/i&gt; spp. and 6.8% MiE, 0.09% ME, and no VME for &lt;i&gt;Enterococcus&lt;/i&gt; spp. Taken together, these data suggest that egAST represents a viable strategy to reduce the prolonged incubation period currently recommended by CLSI and select automated AST manufacturer guidelines without incurring any additional costs while simultaneously maintaining high concordance with reference standard methods.IMPORTANCETraditional antimicrobial-susceptibility testing (AST) methods typically span several days from the time of organism isolation. The majority of this time is spent waiting for a cultured isolate to incubate up to 1 day prior to AST. There exists an unmet need to provide more rapid AST as various rapid methods have been shown to reduce exposure to broad-spectrum antibiotics that select for antimicrobial resistance, shorten hospital stays, and improve clinical outcomes. Simultaneously, there is a need to ensure that rapid AST approac","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0023625"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345138/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144528222","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":"Percentage of culture confirmation and melting curve analysis reveals false-positive <i>Campylobacter</i> detection in a molecular syndromic panel.","authors":"Annie Sanchez, Michael Rauch, Sherry Buechner, Brittney Jung-Hynes, Eric Beck, Allen Bateman","doi":"10.1128/jcm.00028-25","DOIUrl":"10.1128/jcm.00028-25","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Surveillance of enteric organisms can identify outbreaks and support control measures. The BIOFIRE FilmArray Gastrointestinal Panel (FilmArray GI) is the most common culture-independent diagnostic test (CIDT) used in Wisconsin to diagnose enteric infections. State clinical laboratories have raised concerns about inaccurate &lt;i&gt;Campylobacter&lt;/i&gt; detection with the FilmArray GI. The Wisconsin State Laboratory of Hygiene (WSLH) evaluated the percentage of positivity and culture confirmations stratified by CIDT for &lt;i&gt;Campylobacter&lt;/i&gt;, &lt;i&gt;Salmonella&lt;/i&gt;, and STEC from 2018 to 2024. We further analyzed the transport time of &lt;i&gt;Campylobacter&lt;/i&gt; specimens and the melt curves of specimens that tested positive by FilmArray GI. &lt;i&gt;Campylobacter&lt;/i&gt;, &lt;i&gt;Salmonella&lt;/i&gt;, and STEC specimens tested on the FilmArray GI had increases in percentage of positivity compared to those tested on other CIDTs. &lt;i&gt;Salmonella&lt;/i&gt; and STEC specimens positive by FilmArray GI, compared to other CIDTs, had no significant culture confirmation differences. However, &lt;i&gt;Campylobacter&lt;/i&gt; specimens positive by FilmArray GI had significantly lower culture confirmations than positive specimens from other CIDTs (62% vs. 78%; &lt;i&gt;P&lt;/i&gt; &lt; 0.05). &lt;i&gt;Campylobacter c&lt;/i&gt;ulture confirmations were lower for FilmArray GI specimens, compared to other CIDTs, regardless of how many days the specimen spent in transport in 2023. Additionally, we found atypical melt curves among specimens positive by FilmArray GI with cultures that did not grow &lt;i&gt;Campylobacter&lt;/i&gt;. Taken together, the higher percentage of positivity, the lower percentage of culture confirmation, and the atypical melt curves suggest that the FilmArray GI might yield false-positive &lt;i&gt;Campylobacter&lt;/i&gt; results. These findings emphasize the importance of public health surveillance to identify potential issues with commercially available diagnostic tests.IMPORTANCEEnteric pathogens cause ~9.4 million illnesses annually in the United States. Clinical laboratories rely on culture-independent diagnostic testing platforms (CIDTs) for rapid and accurate diagnosis of enteric pathogens. &lt;i&gt;Campylobacter&lt;/i&gt;, &lt;i&gt;Salmonella&lt;/i&gt;, and Shiga-toxin-producing &lt;i&gt;Escherichia coli&lt;/i&gt; (STEC) are three of the most identified enteric bacterial infections in Wisconsin. The BioFire FilmArray Gastrointestinal panel (FilmArray GI) is currently the most common CIDT used by Wisconsin clinical laboratories to diagnose &lt;i&gt;Campylobacter&lt;/i&gt;, &lt;i&gt;Salmonella&lt;/i&gt;, and STEC infections. However, the FilmArray GI has had notable issues and recalls in the past. Here, we used public health surveillance data to assess platform performance for these organisms. We analyzed percent positivity and culture confirmations based on testing platforms. Through our analysis, we identified potential false-positive &lt;i&gt;Campylobacter&lt;/i&gt; results from the FilmArray GI, which were associated with atypical melt curve profiles on the BioFire platform. Public health surveillance can he","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0002825"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345244/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144626430","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 selective culture media for efficient isolation of <i>Avibacterium paragallinarum</i> from chickens.","authors":"Mariela E Srednik, Mostafa M S Shelkamy, Amro Hashish, Nubia R De Macedo, Yuko Sato, Mohamed M El-Gazzar, Orhan Sahin, Qijing Zhang","doi":"10.1128/jcm.00311-25","DOIUrl":"10.1128/jcm.00311-25","url":null,"abstract":"<p><p>Infectious coryza (IC) caused by <i>Avibacterium paragallinarum</i> (AvP) is an upper respiratory disease in chickens and incurs a significant economic impact on laying hens. Control of IC requires reliable bacterial isolation, but AvP is a fastidious bacterium, and the currently used methods yield poor isolation. To address this need, we developed selective media for the efficient growth and isolation of AvP. Several basal media supplemented with various growth factors were explored, and Mueller Hinton agar (MHA) supplemented with fetal bovine serum (FBS) plus NAD yielded optimal AvP growth, eliminating the need for nurse bacteria. This medium (named MSN) was further supplemented with vancomycin and crystal violet to produce two selective media, named MSNV and MSCV, respectively, to inhibit commensal Gram-positive bacteria that reside in the upper respiratory tract of chickens. MSNV and MSCV were compared with the standard isolation methods used in veterinary diagnostic laboratories for AvP isolation using samples from IC-positive and IC-negative flocks verified by clinical observation and AvP-specific RT-PCR. The two selective media significantly increased the isolation of AvP from clinical cases (59.4% for MSNV and 46.9% for MSCV) compared to the conventional method (37.7%). In addition, MSNV and MSCV successfully isolated the recently discovered non-pathogenic AvP variants, which were difficult to obtain using the standard methods. Notably, both media drastically reduced background bacteria and improved the purity of AvP isolates. These results demonstrate the efficacy and usefulness of the selective media for AvP isolation, which will enhance IC diagnosis, antimicrobial susceptibility testing, and vaccine development.IMPORTANCEInfectious Coryza is an economically important disease in poultry, resulting in poor growth and a significant reduction in egg production. The causative agent (AvP) of the disease is difficult to isolate from clinical samples and often requires a nurse bacteria to provide essential nutrients for optimal growth. In addition, the currently used methods for isolation are inefficient and often result in contamination by residential bacterial species. The selective media (MSNV and MSCV) developed in this study solve these problems by eliminating the need for nurse bacteria and by inhibiting Gram-positive bacteria normally present in the respiratory tract of chickens. The media not only increase isolation efficiency but also improve the purity of the isolates. These advancements will facilitate IC diagnosis and the development of vaccines for effective control of this major poultry pathogen.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0031125"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345139/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144659365","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 and validation of next-generation sequencing-based clinical test for triazole resistance prediction in <i>Aspergillus fumigatus</i>.","authors":"J R Caldera, Ashley Dayo, Nathan Wiederhold, Shangxin Yang","doi":"10.1128/jcm.00291-25","DOIUrl":"10.1128/jcm.00291-25","url":null,"abstract":"<p><p>The rising rates of triazole drug resistance in <i>Aspergillus fumigatus</i> have placed greater reliance on antifungal susceptibility testing (AFST) to guide therapeutic management, particularly in medically complicated patients. Current methods, however, utilize conventional phenotypic assays that may pose significant challenges in performance, result interpretation, and time to reporting. Herein, we developed and validated a next-generation sequencing-based clinical test to predict the AFST phenotype of <i>A. fumigatus</i> against voriconazole, posaconazole, isavuconazole, and itraconazole using the <i>cyp51A</i> genotype as a marker for susceptibility. We sequenced 109 isolates comprising reference and clinical samples from the University of California, Los Angeles, Clinical Microbiology Lab, University of Texas Health Science Center at San Antonio Fungus Testing Laboratory, and the Centers for Disease Control and Prevention and Food and Drug Administration Antimicrobial Resistance Isolate Bank. Additionally, we integrated data from 14 previously published studies to produce comprehensive and robust interpretive criteria for the assay. Despite the complex association between <i>cyp51A</i> mutations and drug resistance, our triazole resistance assay produced a remarkable negative percent agreement (specificity) of ≥95%, thus offering presumptive, yet clinically actionable identification of triazole-wild-type isolates. Clinically, this allows for the rapid discrimination between acquired microbiological resistance in <i>A. fumigatus</i> and observed clinical resistance due to patients' declining health to help guide the most effective therapeutic management.IMPORTANCEThe rising rates of antifungal resistance have been expressed by many as \"the silent pandemic,\" profoundly reshaping the landscape of fungal disease management. Innovations in clinical mycology, however, have remained limited, particularly in comparison to the significant advances seen in the greater field of microbiology. Here, we sought to capitalize upon the expanding utility of next-generation sequencing to address a gap in clinical mycology diagnostics and antifungal susceptibility testing. We developed a whole-genome sequencing protocol to evaluate <i>Aspergillus fumigatus cyp51A</i> genotype to predict phenotypic susceptibility to triazole drugs. Our triazole resistance assay offers clinically actionable identification of triazole-wild-type isolates of <i>A. fumigatus</i> in a much more expeditious timeline than traditional phenotypic susceptibility testing.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0029125"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345187/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144540376","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":"Perfect vs practical: utilizing hematology thin smears for the diagnosis of <i>Plasmodium</i> and <i>Babesia</i>.","authors":"Neil Anderson","doi":"10.1128/jcm.00644-25","DOIUrl":"10.1128/jcm.00644-25","url":null,"abstract":"<p><p>In the manuscript entitled \"Hematology Thin Smears Perform Equally to Parasitology Thick and Thin Blood Smears for the Diagnosis of <i>Plasmodium</i> and <i>Babesia</i> in a Low-Prevalence Setting,\" the investigators demonstrate that traditional hematology smears (HS) review can have comparable performance to formal blood parasite smears (PS) review. The authors quote a 93.3% sensitivity in comparison to thick smears. This increases to 100% considering only new diagnoses. The implications are that HS can be utilized as a practical and rapid alternative to PS. For clinical laboratories that are limited to only HS capabilities, these findings suggest that they may be able to be less reliant on reference laboratory PS testing, allowing for more rapid diagnosis and management. Even in clinical laboratories that offer both tests, the findings suggest a reassessment of how both HS and PS fit into the diagnostic process for blood parasites. Familiarity with the pros and cons of both approaches allows them to be employed in a way that maximizes patient care impact.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0064425"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345171/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144608540","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}