BACKGROUND:One of the most successful and widely-distributed hospital-associated lineages of MRSA is clonal complex 5 (CC5). These strains are known for widespread antibiotic resistance but less severe disease than CA-MRSA counterparts. Recently, CC5 descendant lineages have appeared globally with hypervirulent properties. Herein, we identify and characterize a rare and novel CC5 MRSA sequence type, ST3390. METHODS:We used whole genome sequencing, alongside phenotypic characterizations, genetic complementation, blood viability- and neutrophil-killing assays, and a murine model of sepsis to study the pathogenic capabilities of ST3390 strains. RESULTS:To date, there have only been 65 recorded instances of infection caused by ST3390 globally, with 36 of those occurring in Tampa (TPA-ST3390). Genomic analysis of strains identified numerous spa-types, with a t010 cluster found only in our strains. Exploration of AMR genes detected the presence of unique hybrid SCCmec types, with ∼90% of Tampa strains possessing components of SCCmecIa, SCCmecIIa, and/or SCCmecVIII. Phenotypically, all ST3390 strains lack the staphyloxanthin pigment, which is mediated by a conserved 6aa in frame deletion within the staphyloxanthin biosynthesis protein CrtN. TPA-ST3390 strains display high levels of cytotoxicity towards human neutrophils compared with other CC5 lineages and are also virulent in animal models of infection. CONCLUSIONS:This is the first study to characterize the pathogenicity and genomic architecture of the rare MRSA lineage ST3390. Our work provides a deeper understanding of the clonal expansion of CC5, and the wider diversification of Staphylococcus aureus isolates within patient populations.

Candida auris is a multidrug-resistant fungal pathogen associated with healthcare outbreaks and high mortality. Its accurate and timely identification is critical for infection prevention, yet conventional culture-based methods are limited by slow growth, morphological ambiguity, and potential misidentification. This multicenter study evaluated the performance of the Simplexa C. auris Direct assay using 2,020 axilla/groin swab specimens collected from six clinical sites compared to culture followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Specimens were tested prospectively (n = 1,930) and retrospectively (n = 90), with demographic data spanning inpatient, intensive care unit, emergency department, and long-term acute care hospital settings. Discordant results were resolved by bi-directional sequencing, targeting the internal transcribed spacer and D1/D2 regions of the 28S ribosomal DNA gene. Compared to culture followed by MALDI-TOF MS identification, the Simplexa assay demonstrated an overall sensitivity of 94.8% and specificity of 98.7%, with a Cohen's kappa of 0.80, indicating strong agreement. Discordance rates were low (1.4%), and diagnostic accuracy was high (98.6%). The limit of detection was 127 CFU/mL for Clade I and 260 CFU/mL for Clade IV. Mean cycle threshold values were significantly lower in culture-positive specimens (25.1, 95% CI, 23.6-26.7) compared to culture-negative ones (33.6, 95% CI, 32.1-35.0; P < 0.0001). Comparative analysis with four lab-developed molecular tests (LDTs) showed low rates of discordance: 0.9% for LDT 1 and 2, 1.5% for LDT 3, and 0% for LDT 4, indicating a high level of agreement between the molecular methods. The Simplexa C. auris Direct assay provides a rapid and reliable alternative to traditional methods, providing results in under 2 h, supporting early detection and containment of C. auris in healthcare settings. IMPORTANCE:Rapid and accurate detection of Candida auris colonization is essential for preventing healthcare-associated outbreaks and reducing mortality. This multicenter evaluation demonstrates that the Simplexa C. auris Direct assay offers a sensitive, specific, and practical alternative to culture-based methods, enabling earlier identification and containment of C. auris. These findings provide strong evidence to support its implementation in routine infection prevention strategies across diverse healthcare settings.

Background:One of the most successful and widely-distributed hospital-associated lineages of MRSA is CC5. These strains are known from widespread antibiotic resistance, but less severe disease than CA-MRSA counterparts. Recently, CC5 descendant lineages have appeared globally with hypervirulent properties. Methods:Herein we use genomic analyses to study the epidemiology of a rare CC5 MRSA sequence type, ST3390, circulating within Tampa General Hospital (TGH). We employ genetic tools alongside in vitro and in vivo models of virulence to study the pathogenic capabilities of strains. Results:To date, there have only been 50 recorded instances of infection caused by ST3390 globally, with 36 of those occurring at TGH. Genomic analysis of strains identified numerous spa-types, with a t010 cluster found only at TGH. Exploration of AMR genes detected the presence of unique hybrid SCCmec types, with ~90% of TGH strains possessing components of SCCmecIa, SCCmecIIa and/or SCCmecVIII. Phenotypically, all ST3390 strains lack the staphyloxanthin pigment, which is mediated by a conserved 6 aa in frame deletion within the staphyloxanthin biosynthesis protein CrtN. ST3390 strains display high levels of cytotoxicity towards human neutrophils compared to other CC5 lineages, with several isolates displaying hypervirulence in animal models of infection. Conclusions:This is the first study to characterize the pathogenicity and genomic architecture of the rare MRSA lineage ST3390. Our work provides a deeper understanding of the clonal expansion of CC5, and the wider diversification of S. aureus isolates within patient populations.

Antimicrobial resistance (AMR) poses a significant global health threat, projected to cause 10 million deaths annually by 2050. Addressing AMR requires a coordinated, multidisciplinary approach encompassing infectious disease (ID) clinicians, pharmacists, microbiologists, infection preventionists, and policymakers. The inaugural AMR Summit, hosted by bioMérieux in collaboration with Tampa General Hospital and the University of South Florida Morsani College of Medicine in November 2024, convened experts from various fields to explore innovative strategies for combating AMR. Key topics discussed included the role of multidisciplinary teams in antimicrobial stewardship programs, advancements in rapid diagnostic tests and antimicrobial susceptibility testing, the application of implementation science in AMR, and the integration of next-generation sequencing in ID diagnostics. The summit underscored the importance of diagnostic innovation, interdisciplinary collaboration, policy, advocacy, and public engagement in advancing efforts against AMR.

ABSTRACT Timely initiation of appropriate antimicrobial therapy is crucial for patients with Gram-negative (GN) blood stream infections. In this study, the performance of VITEK REVEAL (bioMérieux, USA), an FDA cleared in vitro diagnostic automated system for antimicrobial susceptibility testing (AST) directly from positive blood culture (BC), was compared to that of Accelerate Pheno (Accelerate Diagnostics, USA), as standard of care method. 128 GN positive BCs were analyzed according to manufacturer recommendations, comparing time to result (TTR) and AST results. The categorical agreement (CA) rate between VITEK REVEAL and Accelerate Pheno was 94.3% and the essential agreement (EA) rate was 96.0%. Very major discrepancies (VMD), major discrepancies (MD), and minor discrepancies (miD) rates between the systems were 7.5%, 0.5%, and 4.1%, respectively. Lastly, we observed a mean sequential TTR (reporting of AST results per antibiotic in real-time) on VITEK REVEAL of 6.1 h (3.0–8.2) and mean final TTR of 7.9 h (6.5–8.2), compared to Accelerate Pheno, with a significantly shorter mean final TTR of 7.1 h (6.8–7.7). Sequential TTR on VITEK REVEAL was significantly shorter for resistant isolates (those with ≥1 ‘Resistant’ antimicrobial-organism combination interpretation, using FDA STIC 2024 or CLSI M100 criteria) compared to susceptible/intermediate ones, with a mean difference of 1.4 h (P < 0.001). Overall, compared to Accelerate Pheno, VITEK REVEAL displayed high %CA and %EA for AST of GN bacteria directly from positive BC. Also, unlike Accelerate Pheno, VITEK REVEAL reports MIC results in real time, allowing an earlier release of actionable information useful for antimicrobial stewardship.IMPORTANCEPrevious studies have compared the performance of the VITEK REVEAL, system for fast antimicrobial susceptibility testing (AST) to conventional, non-rapid microbiology methods. To evaluate how the VITEK REVEAL correlates to similarly available fast, direct-from-blood culture AST technologies, this study aimed to compare it to the Accelerate Pheno system, as standard of care method. High categorical agreement (94.3%) and essential agreement (96.0%) between the two systems were observed, underscoring their reliability. The VITEK REVEAL has the advantage of real-time AST reporting, unlike the Accelerate Pheno. Ultimately, this study supports the need for and continued optimization of fast diagnostic technologies for AST, contributing to the goal of advancing antimicrobial stewardship-focused treatment strategies and improving the prognosis of patients with bloodstream infections.

Background:Colonization with Staphylococcus aureus is a risk factor for subsequent infection. Decolonization with the topical antibiotic mupirocin is effective and reduces the risk of subsequent S. aureus infection for both methicillin-sensitive and methicillin-resistant (MRSA) strains but may select for mupirocin-resistant isolates. Methods:We characterized oxacillin and mupirocin susceptibility amongst 384 S. aureus strains isolated from clinical samples isolated in 2017-2023 in Tampa, Florida, spanning strains collected before and after the onset of the coronavirus disease 2019 (COVID-19) pandemic. Whole genome sequencing of bacterial isolates was conducted in parallel and correlated with drug susceptibility profiles. Results:Mupirocin resistance (MupR) was nearly exclusively present in MRSA strains (103/106, 97.1% of MupR; 103/299, 34.4% of MRSA). Although our hospital protocol for decolonization shifted to povidone iodine in the post-COVID period, the overall prevalence of MupR did not change in pre-COVID and post-COVID samples (28.9% vs 26%). Genotype correlated with antibiotic susceptibility with low-level MupR, linked to mutations in ileS and high-level MupR, linked to the presence of mupA. Genome analysis revealed that most MupR strains fell into 3 sequence types (ST) falling into 2 major clonal complexes (CC): CC8 ST8 (including community-associated MRSA strains USA300 and USA500), CC5 ST5 (associated with healthcare-associated MRSA such as USA100), and CC5 ST3390. ST3390 isolates had the highest prevalence of MupR (30/36 83%; high-level MupR 20/36 55.6%; low-level MupR 10/36 27.8%). Conclusions:Mupirocin resistance was prevalent in our hospital MRSA strains. We also found evidence for emergence and persistence of ST3390 MRSA-MupR strains in Florida.

The LIAISON PLEX Respiratory Flex Assay is a customizable syndromic molecular assay for the qualitative detection of nucleic acids from 14 viral and 5 bacterial respiratory pathogens in nasopharyngeal swabs (NPS). This study evaluated its clinical performance in comparison to established FDA-cleared and standard-of-care molecular diagnostic platforms. A total of 2,099 NPS specimens were prospectively and retrospectively collected. Positive Percent Agreement (PPA), Negative Percent Agreement (NPA), and diagnostic accuracy were calculated across 19 targets. The assay was benchmarked against SOC assays, including Cepheid Xpert Xpress CoV-2/Flu/RSV Plus, Abbott Alinity m Resp-4-Plex, and BioFire RP 2.1 panel. Agreement metrics, kappa coefficients, and discordance analysis were used to assess the comparative performance. For bacterial targets, PPA ranged from 92.3% to 100%, with NPA exceeding 99%, and diagnostic accuracy above 99.7%. Viral targets showed PPA from 90.3% to 100% and NPA from 95.8% to 100%. Adenovirus, Influenza A, and Influenza B achieved 100% PPA. RSV and SARS-CoV-2 demonstrated diagnostic accuracies of 99.7% and 99.0%, respectively. Agreement with SOC assays was high (PPA up to 97.1%; kappa 0.81-0.97). Discordance analyses identified differences in pathogen detection, with the Respiratory Flex Assay detecting analytes missed by SOC assays. The LIAISON PLEX Respiratory Flex Assay demonstrated high accuracy, reliability, and strong diagnostic agreement with SOC assays. Its robust performance across respiratory pathogens supports its use as a comprehensive diagnostic tool for respiratory infection management. IMPORTANCE:Fast and accurate detection of respiratory pathogens helps guide treatment, lower healthcare costs, and support disease monitoring. The LIAISON PLEX Respiratory Flex Assay was evaluated for respiratory pathogen detection by comparing its performance to FDA-cleared molecular respiratory pathogen detection methods, PCR followed by bi-directional sequencing (PCR/BDS) for bacterial pathogens, and standard-of-care (SOC) molecular respiratory assays, including targeted and multiplex syndromic PCR panels. The study assessed its ability to identify pathogens associated with respiratory tract infections, measure agreement with SOC assays, and determine its accuracy in clinical settings. The findings provide data on the assay's diagnostic performance, testing flexibility, and applicability in laboratory workflows.

Background: BK virus (BKV) is the main cause of polyomavirus‐associated nephropathy in kidney transplant patients and hemorrhagic cystitis in bone marrow recipients. BKV quantitation by PCR is crucial in diagnostic and therapeutic management of transplant patients infected with BKV. We evaluated the performance of the Qiagen BKV ASR for the quantification of BKV on the NeuMoDx™ 96 System and compared the results to our standard of care (SOC) test, the Diasorin BKV ASR on the Liaison® MDX. Methods: The analytical performance was assessed using commercially available BKV Panels that meet the 1st WHO International Standards for BKV nucleic acid amplification techniques. The clinical performance was evaluated using 204 residual plasma and urine samples previously identified with the SOC assay. Results: The assay exhibited a strong linear correlation (R² = 0.9985) with the reference panel and an excellent analytical accuracy (R² = 0.9976). The LoD was determined to be 50 IU/mL with remarkable precision within and between days (SDEV 0.00—0.57 and 0.05—0.31, respectively). Of the 204 samples, only 10 (4.9 %) were discordant (PPA = 92.37 %; NPA = 100 %). Although the Qiagen BKV ASR quantified BKV DNA at an average of 0.48 Log IU/mL lower than the SOC, it showed a strong concordance to the SOC results. Compared to the SOC, the Qiagen BKV ASR had a more automated workflow, with less hands-on time, leading to quicker turnaround time. Conclusion: The Qiagen BKV ASR is specific, sensitive, and accurate in quantifying BKV in plasma and urine specimens on the fully automated NeuMoDx™ 96 System.

Abstract Background The adaptive power of Staphylococcus aureus enables it to cause a wide variety of infections and readily develop resistance to antibiotics. It does this through extensive variability in its genomic repertoire, not only via laterally acquired elements but also through core genome evolution. Methods Herein we use whole genome sequencing to study the genomic epidemiology of S. aureus strains circulating within Tampa General Hospital (TGH). Results While characterizing the genetics of mupirocin resistance, we identified a rare ST belonging to hospital-associated clonal complex 5. The elevated rate of mupirocin resistance in this ST, ST3390 (83.2% total, high-level resistance 55.5% and low-level resistance 27.7%), was particularly striking. To date, there have only been 6 recorded instances of this lineage globally, whilst this work identified 36 individual strains from TGH alone. Genome analysis of our strains identified numerous completely conserved non-synonymous mutations in genes relating to DNA damage response, metabolism, adhesion/biofilm formation, iron acquisition and peptidoglycan biosynthesis compared to CC5 isolates. Additionally, exploration of AMR genes detected the presence of a unique dual SCCmec type, with two-thirds of strains possessing components of both SCCmecIa and SCCmecIIa. Phenotypically, all ST3390 strains lack the characteristic staphyloxanthin pigment that gives S. aureus its name. Additionally, ST3390 isolates display high levels of cytotoxicity towards human neutrophils compared to other CC5 strains. Conclusion Collectively, this work provides a deeper understanding of the genetic variability of S. aureus in clinical settings, leading to a better comprehension of in-hospital bacterial population dynamics and the evolution and spread of contemporary isolates. Disclosures All Authors: No reported disclosures

Abstract Background Mupirocin (Mup) is a topical drug used to decolonize carriers of Staphylococcus aureus (SA) to prevent later SA infection. A spot survey of SA isolates at Tampa General Hospital (TGH) revealed a prevalence in Mup resistance of ∼40%, leading to a change of decolonization protocol from mupirocin to povidone iodine in January 2020, just prior to the COVID pandemic. We determined the prevalence of Mup resistance in a random sampling of clinical isolates before and after change in decolonization protocol. Methods A total of 211 PreCOVID (before 2020) and 173 PostCOVID (after 2021) samples from various sources (blood, bone, eye, fluid, respiratory, urine, and wound) were tested with minimum inhibitory concentration for Mup resistance using literature values for susceptible, low resistance, and high resistance. Oxacillin sensitivity, to verify MRSA isolates, was tested in parallel. Results were analyzed via χ² for significance (p=0.05). Results Comparison of Mup resistance levels between the Pre- and PostCOVID time points showed no significant difference (p=0.460). PreCOVID: 71.1% susceptible, 8.1% low resistance, 20.9% high resistance. PostCOVID: 74.0% susceptible, 9.8% low resistance, and 16.2% high resistance. MRSA status correlated with Mup resistance in both Pre- (p< 0.001) and PostCOVID samples (p=0.001). Blood vs Non-Blood samples differed in Mup resistance in the PreCOVID period (20.7% blood vs 34.1% non-blood, p=0.037), whereas samples in the PostCOVID period showed no significant difference in Mup resistance (22.8% blood vs 28.7% non-blood, p=0.375). Notably, 46.2% PreCOVID Respiratory were resistant to Mup (p=0.016). 26.3% PostCOVID Respiratory were resistant to Mup (p=0.651). Conclusion Change in decolonization protocol did not influence the overall prevalence of Mup resistance. Respiratory samples made up a significant proportion of the resistant samples in the PreCOVID time point, suggesting differences in prevalence of Mup resistance in different clinical sites. Further molecular epidemiological studies are needed to determine if specific sites of infection or specific patient groups are more likely to be colonized with Mup resistant SA isolates. Disclosures All Authors: No reported disclosures

Acute gastroenteritis (AGE) is a leading cause of morbidity and mortality worldwide across all age groups that disproportionally affects young children in low- and middle-income countries and immunocompromised patients in high-income countries. Regional outbreaks of AGE are typically detected by traditional microbiological detection methods that target limited organisms and are associated with low sensitivity and lengthy time-to-results. Combined, these may result in repeat testing, imprecise or delayed treatment, and delayed recognition of outbreaks. We conducted a multi-site prospective study comparing the BioCode Gastrointestinal Pathogen Panel (BioCode GPP) for the detection of 17 common bacterial, viral, and protozoan causes of gastroenteritis with reference methods, including stool culture, enzyme immunoassays, pathogen-specific PCR assays, and sequencing. One thousand five hundred fifty-eight residual, de-identified stool samples (unpreserved stool and stool in Cary-Blair transport medium) were enrolled and tested for 11 bacterial, 3 viral, and 3 protozoan pathogens. BioCode GPP and reference methods were positive for 392 (25.2%) and 283 (18.2%) samples, respectively (P < 0.0001). In this study, the BioCode GPP and reference methods detected 69 and 65 specimens positive for Clostridioides difficile, 51 and 48 for enteroaggregative Escherichia coli, 33 and 27 for enterotoxigenic E. coli, 50 and 47 for norovirus GI/GII, and 30 and 22 for rotavirus A, respectively. The BioCode GPP showed good positive and negative agreements for each pathogen ranging from 89.5% to 100%, with overall sensitivity and specificity of 96.1% and 99.7%, post adjudication. The BioCode GPP detected >1 pathogens in 49 samples, representing 12.5% of the total 392 positive specimens. IMPORTANCE:This study highlights performance of a novel technology for timely and accurate detection and differentiation of 17 common bacterial, viral, and protozoan causes of gastroenteritis. Utilizing molecular tests such as the BioCode Gastrointestinal Pathogen Panel may improve the detection of gastrointestinal pathogens and provide actionable results, particularly for patient populations at most risk.

Spa-typing is a major genetic tool used to distinguish between Staphylococcus aureus strains. Interestingly, although rare, similar to 1%-2 of isolates are considered Spa-non-typeable. Herein, we present the draft genome sequence of just such a strain, S. aureus TGH1097, a USA300 isolate from a complex bacteremia infection.

BACKGROUND:Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are routinely tested and reported; however, Trichomonas vaginalis (TV) is the most common sexually transmitted infection (STI) in the United States and the prevalence of Mycoplasma genitalium (MG) infections is likely higher than estimated. We examined the clinical performance of the Alinity m STI assay for detection and surveillance of CT/NG/TV/MG in urine specimens from patients at a large academic medical center. METHODS:Urine specimen from 198 patients was tested in this evaluation. Alinity m STI and Aptima Combo 2 CT/NG and TV assay (Panther System) results were compared, with discrepant results run on the cobas 6800 CT/NG, TV/MG assays. Analyzer turnaround times, time from loading the specimen on the analyzer to results reporting, were determined for Alinity m and Panther systems. RESULTS:Overall percent agreements of the Alinity m in comparison with the Aptima and cobas assays for CT, NG, TV, and MG were 99.5% (97.2%, 99.9%), 99.5% (97.2%, 99.9%), 98.4% (95.5%, 99.5%), and 86.4% (66.7%, 95.3), respectively. There were 5 discrepant samples (CT, 1; NG, 1; TV, 3) between the Alinity m and the Aptima assays, and 3 MG discrepant samples between the Alinity m STI and cobas 6800. Two of the 5 Aptima and Alinity m discrepant samples were resolved as they yielded similar results on both Alinity m and cobas 6800. TV and MG infections comprised 54% of the positive samples and were more often asymptomatic than CT and NG infections. Analyzer turnaround time was 3 hours 25 minutes for the Aptima CT/NG, 3 hours 25 minutes for Aptima TV, and 1 hour 55 minutes for Alinity m STI assay. CONCLUSIONS:The Alinity m STI assay allows for fast and simultaneous detection of the 4 major STI pathogens, which can facilitate surveillance and provide accurate results to help clinicians diagnose for initiation of appropriate treatment.

West Nile virus (WNV) is an arbovirus spread primarily by Culex mosquitoes, with humans being a dead-end host. WNV was introduced to Florida in 2001, with 467 confirmed cases since. It is estimated that 80 percent of cases are asymptomatic, with mild cases presenting as a non-specific flu-like illness. Currently, detection of WNV in humans occurs primarily in healthcare settings via RT-PCR or CSF IgM when patients present with severe manifestations of disease including fever, meningitis, encephalitis, or acute flaccid paralysis. Given the short window of detectable viremia and requirement for CSF sampling, most WNV infections never receive an official diagnosis. This study utilized enzyme-linked immunosorbent assay (ELISA) to detect WNV IgG antibodies in 250 patient serum and plasma samples collected at Tampa General Hospital during 2020 and 2021. Plaque reduction neutralization tests were used to confirm ELISA results. Out of the 250 patients included in this study, 18.8% of them were IgG positive, consistent with previous WNV exposure. There was no relationship between WNV exposure and age or sex.

The monkeypox (Mpox) virus has raised significant concerns given its recent spread with an increasing number of confirmed cases worldwide. In this study, we evaluated the performance of a laboratory developed test (LDT) using BioGX Xfree hMPXV/OPXV reagents for the qualitative detection of non-variola Orthopoxviruses and Mpox virus DNA, in swabs from human pustular or vesicular rash specimens. Analytical and clinical testing analysis were carried out on two different platforms: the BD MAX™ System (BD Diagnostics) and the new pixl.16 Real-Time PCR Platform (BioGX), using a synthetic Mpox virus DNA (ATCC VR-3270SD) and residual clinical samples previously identified with an EUA approved Mpox real-time PCR assay. In the end, the Xfree hMPXV/OPXV LDT proved to be a sensitive, specific, and reproducible test for the detection of Mpox on both platforms evaluated with the pixl.16 having an advantage of a small footprint and providing faster TAT facilitated by an extraction-free workflow.

During May-July 2023, a cluster of 7 patients at local hospitals in Florida, USA, received a diagnosis of Plasmodium vivax malaria. Whole-genome sequencing of the organism from 4 patients and phylogenetic analysis with worldwide representative P. vivax genomes indicated probable single parasite introduction from Central/South America.

The secreted proteases of Staphylococcus aureus have been shown to be critical during infection. Here, we present the draft genome sequence of S. aureus TGH337, a hyper-proteolytic USA300 strain isolated from human urine.

Early detection of microbial pathogens causing respiratory tract infection plays a crucial role in clinical management. The BioCode Respiratory Pathogen Panel (BioCode RPP) utilizes reverse transcriptase PCR (RT-PCR) in combination with barcoded magnetic beads to amplify, detect, and identify respiratory pathogens. This panel qualitatively detects and identifies 14 viruses, including influenza virus A with H1 pdm09, H1, and H3 subtyping; influenza B; respiratory syncytial virus (RSV); human metapneumovirus; parainfluenza virus 1; parainfluenza virus 2; parainfluenza virus 3; parainfluenza virus 4; coronavirus (229E, NL63, OC43, and HKU1); adenovirus; and human rhinovirus/enterovirus, and 3 bacteria, including Chlamydia pneumoniae, Mycoplasma pneumoniae, and Bordetella pertussis. Reproducibility, which was assessed with contrived specimens containing 12 targets at 3 clinical sites, with 2 operators at each site for 5 days, was 99.4% for Flu A H3 and Flu B, 98.9% for RSV, and 100% for the remaining 9 targets assayed. A multicenter clinical trial evaluated the performance of the BioCode RPP with 2,647 nasopharyngeal swab specimens from 5 geographically distinct sites and revealed comparable performance between the BioCode RPP and FilmArray Respiratory Panel (FA-RP). Specifically, the positive percent agreements (PPAs) for various pathogens ranged between 80.8% and 100% compared with the FA-RP (1.7 and 2.0). Negative percent agreement ranged from 98.4% to 100% for BioCode RPP. The BioCode RPP also offers scalable automated testing capability of up to 96 specimens in a single run with total sample-to-result time under 5 h. The invalid rate of the BioCode RPP on initial testing was 1.0% (26/2,649).IMPORTANCE Early detection of microbial pathogens causing respiratory tract infection plays a crucial role in clinical management. The BioCode Respiratory Pathogen Panel (BioCode RPP) is a high-throughput test that utilizes RT-PCR in combination with barcoded magnetic beads to amplify, detect, and identify 17 respiratory pathogens, including 14 viruses and 3 bacteria. This study summarizes data generated from a multicenter clinical trial evaluating the performance of the BioCode RPP on 2,647 nasopharyngeal swab specimens from five geographically distinct sites.

Background: Cytomegalovirus (CMV) infection has a major negative impact on transplantation and is associated with increased morbidity and mortality in this patient population. Quantitation of CMV infections using a mo-lecular test is the preferred method for monitoring patients post-transplant. For this analysis, we compared the Aptima CMV Quant Assay (Aptima CMV) on the Panther system to the ELITech MGB Alert (R) CMV 3.0 ASR (MGB CMV) run on the ELITe InGenius (R). Methods: The analytical performance of the assay was assessed using commercially available CMV reference panels that meet the 1st WHO International Standard for Human Cytomegalovirus for nucleic acid amplification techniques. The clinical performance of the assay was determined using 249 plasma and non-plasma samples. Results: The 95% LOD of the Aptima assay was determined to be 50 IU/mL and 200 IU/mL for the MGB CMV assay. A strong linear correlation with the reference panel (R2 = 0.9945), excellent reproducibility, and accuracy (R2 = 0.986) over the detection range of the assay was observed. Of the 249 clinical samples tested, only 17 (6.8%) yielded discordant results which were at or near the lower limit of quantification of the assays. Although the Aptima CMV assay demonstrated excellent concordance of qualitative results to the MGB CMV assay for all samples, the MGB CMV quantified CMV DNA at an average of 0.5 Log IU/mL higher than Aptima CMV. Conclusion: The Aptima CMV assay is both sensitive and accurate in quantifying CMV in both plasma and non-plasma specimens on the fully automated Panther system.