Mariela E Srednik, Mostafa M S Shelkamy, Amro Hashish, Nubia R De Macedo, Yuko Sato, Mohamed M El-Gazzar, Orhan Sahin, Qijing Zhang
{"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}
J R Caldera, Ashley Dayo, Nathan Wiederhold, Shangxin Yang
{"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}
Sohei Harada, Kohji Komori, Kenya Yukawa, Brian Hayama, Kazumi Takehana, Kageto Yamada, Asako Doi, Tomoo Saga, Masakazu Sasaki, Yoshiro Hadano, Masahiro Suzuki, Kyoko Yokota, Jun Suzuki, Koki Kikuchi, Yohei Doi, Kazuhiro Tateda
{"title":"Genomic epidemiology and antimicrobial resistance of <i>Corynebacterium macclintockiae</i>, the predominant species of human pathogens within the <i>Corynebacterium jeikeium</i> complex.","authors":"Sohei Harada, Kohji Komori, Kenya Yukawa, Brian Hayama, Kazumi Takehana, Kageto Yamada, Asako Doi, Tomoo Saga, Masakazu Sasaki, Yoshiro Hadano, Masahiro Suzuki, Kyoko Yokota, Jun Suzuki, Koki Kikuchi, Yohei Doi, Kazuhiro Tateda","doi":"10.1128/jcm.00500-25","DOIUrl":"10.1128/jcm.00500-25","url":null,"abstract":"<p><p>Genomic characteristics and optimal treatment of <i>Corynebacterium jeikeium</i> remain largely unknown. We collected clinical information and performed whole-genome sequencing analysis of the causative strains of six cases of <i>C. jeikeium</i> infection at a single hospital over a 9-year period. Additionally, whole-genome sequencing analysis was performed on 33 <i>C</i>. <i>jeikeium</i> strains from cases of bloodstream infection at eight hospitals. Antimicrobial susceptibility testing was performed, and the results were compared to the resistance genes identified. Publicly available genome data of strains of <i>C. jeikeium</i> complex, consisting of <i>C. jeikeium sensu stricto</i>, <i>Corynebacterium macclintockiae</i>, and <i>Corynebacterium evansiae</i>, worldwide, were combined with the data from this study to determine the distribution of genomic species. In the single-center study, cases of prosthetic osteoarticular infection, postoperative intra-abdominal infection, and catheter-related bloodstream infection were identified, and the causative strains were genomically identified as <i>C. macclintockiae</i>. All but one isolate (32/33, 97.0%) in the eight-center study identified as <i>C. jeikeium</i> by matrix-assisted laser desorption ionization-time of flight mass spectrometry were also genomically identified as <i>C. macclintockiae</i>. Nosocomial transmission was suggested in three strain pairs by core-genome single nucleotide polymorphism analysis. <i>C. macclintockiae</i> strains were generally multidrug-resistant, but all anti-methicillin-resistant <i>Staphylococcus aureus</i> agents, including teicoplanin, had favorable activity, and the strains without the <i>tet</i>(W) gene (22/38, 57.9%) were susceptible to tetracyclines. Genome analysis of 66 <i>C</i>. <i>jeikeium</i> complex strains collected worldwide, consisting mainly of clinical strains, re-identified 51 strains (77.3%) as <i>C. macclintockiae</i>. This study demonstrates that <i>C. macclintockiae</i> is the major genomic species of the <i>C. jeikeium</i> complex causing human infections.IMPORTANCERecent widespread use of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has facilitated the identification of <i>Corynebacterium</i> spp. in microbiology laboratories, thereby raising awareness of the clinical importance of these organisms. Nevertheless, the accumulation of information on genomic characteristics of <i>Corynebacterium jeikeium</i> has been significantly limited compared to other pathogenic organisms thus far. In this study, we analyzed causative strains of infections identified as <i>C. jeikeium</i> by MALDI-TOF MS, collected from multiple institutions throughout Japan, and found that most of these strains were genomically identified as <i>Corynebacterium macclintockiae</i>, a species that has been newly described recently. Collection of clinical information on selected cases showed that <i>C. macclintockiae</i> ","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0050025"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345161/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144698695","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":"An ELISA-like sensitive and visual detection system targeting <i>Yersinia pestis</i> based on CRISPR/Cas12a and DNAzyme.","authors":"Yingqing Mao, Ruichen Lv, Hao Shao, Yong Zhao, Junhu Wang, Qiong Chen, Haiming Yi, Yixin Ge, Hongming Wang, Yuexi Li, Yong Qi","doi":"10.1128/jcm.00274-25","DOIUrl":"10.1128/jcm.00274-25","url":null,"abstract":"<p><p><i>Yersinia pestis</i> is the causative agent of plague, a human disease with potentially devastating consequences. Here, we developed an enzyme-linked immunosorbent assay-like visual detection method based on clustered regularly interspaced short palindromic repeats (CRISPR) detection and DNAzyme for the cost-effective and highly sensitive detection of <i>Y. pestis</i>. A novel specific gene sequence (CH57_3927) was screened for the detection target of <i>Y. pestis</i>. The recombinase-aided amplification (RAA) assay, CRISPR/Cas12a detection assay, and G-quadruplex (G4) DNAzyme-based color development assay were separately established and optimized. These three optimized assays were integrated into an advanced ELISA-like visual detection method-RAA-CRISPR/Cas12a-DNAzyme (RCCD)-by further optimization of their components to improve the compatibility between them. The amplified target sequence binds to crRNA and activates the Cas12a nucleases for trans-cleave G4. As a result, the cleaved G4 is unable to bind with hemin to exert peroxidase activity, thus impeding the catalysis of the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS<sup>2-</sup>) colorimetric reaction. Consequently, negative samples exhibit a dark green coloration, while the positive products appear nearly colorless, facilitating visual differentiation with the naked eye. In addition, the RCCD detection platform effectively distinguished <i>Y. pestis</i> from all other closely related species, with a detection limit of 1 copy/reaction. Evaluated using <i>Y. pestis</i> DNA-spiked blood samples and uninfected samples, both sensitivity and specificity were 100%. The method shows significant potential for detecting targets in clinical samples and is well-suited for use in resource-limited environments. It offers advantages such as visual detection, batch detection, and low cost.IMPORTANCEWe utilized Mauve software to screen <i>Yersinia pestis</i> specific genes and integrated CRISPR-Cas12a, RAA amplification, and G-quadruplex DNAzyme technology to establish an advanced ELISA-like visual detection method. The visual detection method offers a more cost-effective alternative compared to the conventional CRISPR detection method that relies on fluorescence-labeled ssDNA reporter or lateral flow (LF) test strips. With only one thermostatic device required, it enhances the convenience of rapid on-site screening of <i>Y. pestis</i> outbreaks, providing effective support for plague detection, prevention, and control within primary medical and health institutions.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0027425"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345247/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144698694","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 BIOBALL and EZ-Accu Shot for accurate quantification as certified quality control reference material.","authors":"Dustin A Baird, James E T Gebo, Anna F Lau","doi":"10.1128/jcm.00520-25","DOIUrl":"10.1128/jcm.00520-25","url":null,"abstract":"","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0052025"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345222/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144540375","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}
Piyash Bhattacharjee, Sonia T Hegde, Ashraful Islam Khan, Imrul Kayes Nabil, Md Naiem Hossain, Tahira Ahmed Rashmi, Mokibul Hassan Afrad, Md Taufiqul Islam, Mohammad Ashraful Amin, Zahid Hasan Khan, Taufiqur Rahman Bhuiyan, Andrew S Azman, Firdausi Qadri
{"title":"Rapid tests as a practical alternative to slide agglutination for the confirmation of <i>V. cholerae</i> O1.","authors":"Piyash Bhattacharjee, Sonia T Hegde, Ashraful Islam Khan, Imrul Kayes Nabil, Md Naiem Hossain, Tahira Ahmed Rashmi, Mokibul Hassan Afrad, Md Taufiqul Islam, Mohammad Ashraful Amin, Zahid Hasan Khan, Taufiqur Rahman Bhuiyan, Andrew S Azman, Firdausi Qadri","doi":"10.1128/jcm.00433-25","DOIUrl":"10.1128/jcm.00433-25","url":null,"abstract":"<p><p>Slide agglutination for serogroup and/or serotype identification is a crucial step for confirming cholera by culture. Rapid diagnostic tests (RDTs) are typically used directly on stool but are not considered sufficient for cholera confirmation. However, they may provide a practical alternative to the slide agglutination step of culture<i>,</i> as they are easy to use, store, and require minimal training. This study evaluates the concordance of <i>Vibrio cholerae</i> O1/O139 detection from presumptive colonies by slide agglutination with RDTs. Patients (≥1 year) with acute watery diarrhea at the icddr,b Dhaka hospital were enrolled. Stool samples were cultured on thiosulfate-citrate-bile salts-sucrose medium, and presumptive colonies were subcultured on Gelatin Agar medium. Isolates were then tested by slide agglutination and four commercial cholera RDTs. From 4 February 2024 through 24 May 2025, 1,638 patients with acute watery diarrhea were enrolled, 1,140 (70%) had presumptive <i>V. cholerae</i> colonies, and 482 (29%) were culture-confirmed as <i>V. cholerae</i> O1. We tested suspected <i>V. cholerae</i> colonies using Cholkit (n=1,140), SD Bioline (n=693), Crystal VC O1/O139 (n=693), and Crystal VC O1 (n=655) RDT kits. RDTs showed near-perfect concordance with slide agglutination. Using slide agglutination as the reference, the sensitivity of the RDT kits ranged from 99.3% to 99.6% and the specificity from 99.3% to 99.7%. RDTs offer a practical and potentially easier alternative to slide agglutination of presumptive <i>V. cholerae</i> colonies within typical cholera culture protocols. This may help to provide a pathway to quick confirmation of outbreaks in settings where lab facilities and reagents may be limited.IMPORTANCEThis study demonstrated that rapid diagnostic tests (RDTs) can effectively replace traditional slide agglutination methods to confirm suspected <i>Vibrio cholerae</i> colonies belonging to the O1 serogroup, which are responsible for the current seventh cholera pandemic. Comparing four RDTs to slide agglutination, we found almost perfect agreement, with sensitivity and specificity exceeding 99%. Since RDTs are cheaper, easier to use and store, and require less technical training, adopting them could significantly speed up cholera outbreak confirmation, especially in areas with limited laboratory resources. This practical shift could lead to faster responses to cholera outbreaks, ultimately improving public health surveillance and disease control globally.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0043325"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345242/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144649608","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}
Valeria Fabre, Yea-Jen Hsu, Karen C Carroll, Aaron M Milstone, Alejandra B Salinas, Lilian M Abbo, Chris Bower, Jennifer Berry, Sarah Boyd, Kathleen O Degnan, Pragya Dhaubhadel, Daniel J Diekema, Marci Dress, Baevin Feeser, Mark Fisher, Cynthia Flynn, Bradley A Ford, Erin B Gettler, Laurel J Glasser, Jessica Howard-Anderson, J Kristie Johnson, Sara M Karaba, Justin J Kim, Alyssa Kubischta, Benjamin M Landrum, Marvin Martinez, Amy J Mathers, Leonard Mermel, Rebekah W Moehring, John C O'Horo, Dana E Pepe, S Sonia Qasba, Barry Rittmann, Evan D Robinson, Guillermo Rodríguez-Nava, Rossana Rosa, Jonathan H Ryder, Jorge L Salinas, Aditya Shah, Gregory M Schrank, Mark Shelly, Emily S Spivak, Kathleen O Stewart, Thomas R Talbot, Trevor C Van Schooneveld, Anastasia Wasylyshyn, Avinash Gadala, Zunaira Virk, Sara E Cosgrove
{"title":"Multicenter evaluation of blood culture contamination and blood cultures practices in US acute care hospitals: time for standardization.","authors":"Valeria Fabre, Yea-Jen Hsu, Karen C Carroll, Aaron M Milstone, Alejandra B Salinas, Lilian M Abbo, Chris Bower, Jennifer Berry, Sarah Boyd, Kathleen O Degnan, Pragya Dhaubhadel, Daniel J Diekema, Marci Dress, Baevin Feeser, Mark Fisher, Cynthia Flynn, Bradley A Ford, Erin B Gettler, Laurel J Glasser, Jessica Howard-Anderson, J Kristie Johnson, Sara M Karaba, Justin J Kim, Alyssa Kubischta, Benjamin M Landrum, Marvin Martinez, Amy J Mathers, Leonard Mermel, Rebekah W Moehring, John C O'Horo, Dana E Pepe, S Sonia Qasba, Barry Rittmann, Evan D Robinson, Guillermo Rodríguez-Nava, Rossana Rosa, Jonathan H Ryder, Jorge L Salinas, Aditya Shah, Gregory M Schrank, Mark Shelly, Emily S Spivak, Kathleen O Stewart, Thomas R Talbot, Trevor C Van Schooneveld, Anastasia Wasylyshyn, Avinash Gadala, Zunaira Virk, Sara E Cosgrove","doi":"10.1128/jcm.00530-25","DOIUrl":"10.1128/jcm.00530-25","url":null,"abstract":"<p><p>Clinical and Laboratory Standards Institute (CLSI) recommends a blood culture contamination (BCC) threshold of <3%, with ≤1% considered optimal. However, there is not a standardized definition of BCC, and the effect of multiple definitions on BCC rates or what definitions laboratories use remain unknown. We surveyed 52 hospitals and analyzed 362,078 blood cultures (BCx) collected 1 September 2019 to 31 August 2021 from 62 intensive care units (ICUs) and 231 wards from 48 of these hospitals. We calculated and compared BCC rates using the College of American Pathologists (CAP) or CLSI criteria (both utilize a limited number of skin commensals to define BCC) and the comprehensive National Healthcare Safety Network (NHSN) commensal list. We characterized factors associated with BCC and related outcomes (central-line associated bloodstream infection [CLABSI] and vancomycin use). BCC, BCx positivity, and single BCx rates were monitored by 100%, 39%, and 21% of hospitals, respectively. Hospitals used CAP (65%), CLSI (17%), and NHSN (17%) criteria to define BCC. Mean BCC rate by CAP (CAP-BCC) was 1.38% for ICUs and 0.96% for wards. BCC rates remained similar by CLSI criteria but increased when using NHSN list. Sharing BCC data outside of the laboratory, measuring additional BCx quality indicators, and limiting central catheter-drawn BCx were associated with lower BCC rates. BCC was associated with higher CLABSI rates in ICUs. This study demonstrated variability in laboratory practices and opportunities to optimize BCx stewardship.IMPORTANCEBlood culture contamination (BCC) is associated with patient harm and unnecessary use of healthcare resources. BCC thresholds have been established; however, multiple BCC definitions exist. There is limited data on how BCC rates differ depending on the BCC definition used, what definitions laboratories most commonly use, or their approach to other blood cultures (BCx) quality indicators such as single rates or BCx positivity. A cross-sectional multicenter survey and analysis of BCx data from intensive care unit and wards revealed that most laboratories did not track single BCx or BCx positivity rates and that there was variability in how BCC was defined. Additionally, BCC rates were influenced by the definition used. BCC was associated with increased central-line associated bloodstream infection rates.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0053025"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345234/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144608538","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}
Elissa M Robbins, Rasa Bertuzis, Ho-Chen Chiu, Lupe Miller, Christopher Noutsios
{"title":"A multicenter study to assess the performance of the point-of-care RT-PCR Cobas SARS-CoV-2 & Influenza A/B nucleic acid test for use on the Cobas Liat system in comparison with centralized assays across healthcare facilities in the United States.","authors":"Elissa M Robbins, Rasa Bertuzis, Ho-Chen Chiu, Lupe Miller, Christopher Noutsios","doi":"10.1128/jcm.01459-24","DOIUrl":"10.1128/jcm.01459-24","url":null,"abstract":"<p><p>Respiratory diseases can share many of the same symptoms, highlighting the need for timely and accurate differentiation to facilitate effective clinical management and reduce transmission. Compared with centralized testing, molecular point-of-care tests (POCTs) can provide a faster time to result. We evaluated the RT-PCR POCT Cobas® SARS-CoV-2 & Influenza A/B qualitative assay for use on the Cobas Liat system (the POC SARS-CoV-2 & Influenza A/B test) in nasal and nasopharyngeal swab samples from 10 diverse healthcare facilities in the United States. A composite comparator design consisting of three centralized tests was used to analyze severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), while performance vs a single centralized test was used for analysis of influenza A and B. Evaluations included performance stratified by sample type (prospective/retrospective and nasal/nasopharyngeal [paired by subject]), collection method (self/healthcare worker-collected [alternated and approximately balanced]), symptom status (symptomatic/asymptomatic), and SARS-CoV-2 vaccination status, as well as assay inclusivity and system ease of use. A total of 2,247 samples were tested. For SARS-CoV-2, the overall percent agreement (OPA) was 98.8% (95% confidence interval [CI]: 97.9, 99.3) in nasal swab samples and 99.0% (95% CI: 98.2, 99.4) in nasopharyngeal swab samples. Regression analysis showed that cycle threshold values from paired nasal and nasopharyngeal swab samples were highly correlated (correlation coefficient 0.83). The OPA was ≥99.5% (sample type dependent) and 100.0% for influenza A and B, respectively. The POC SARS-CoV-2 & Influenza A/B test was easy to use. These results support the use of the POCT in various sample types and by various operators in the intended-use setting.</p><p><strong>Importance: </strong>This study highlights the benefits of RT-PCR point-of-care tests, namely comparable performance to centralized testing in multiple sample types and ease of use. Utilizing assays such as the POC Cobas SARS-CoV-2 & Influenza A/B test may improve the timely differentiation of respiratory diseases that share similar symptoms.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0145924"},"PeriodicalIF":5.4,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12345166/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144540374","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 <i>Staphylococcus aureus</i> complex: implications for the clinical microbiology laboratory.","authors":"Austin Yan, Julianne V Kus, Nadia Sant","doi":"10.1128/jcm.01276-24","DOIUrl":"10.1128/jcm.01276-24","url":null,"abstract":"<p><p>In the last 15 years, advances in diagnostic microbiology have enabled more detailed characterization of human bacterial pathogens. These changes have led to the description of new species within the <i>Staphylococcus aureus</i> complex, including <i>S. aureus</i> subspecies <i>anaerobius</i>, <i>Staphylococcus argenteus</i>, and <i>Staphylococcus schweitzeri</i>. In this minireview, we discuss the history, epidemiology, microbiology, genomics, and clinical significance of the <i>S. aureus</i> complex and provide a guide for laboratories to reliably detect and differentiate these novel species.</p>","PeriodicalId":15511,"journal":{"name":"Journal of Clinical Microbiology","volume":" ","pages":"e0127624"},"PeriodicalIF":6.1,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12239731/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144215992","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}