Drinking water treatment aims to stabilize bacterial communities in drinking water and to remove human pathogens from the system. Legionella pneumophila , Pseudomonas aeruginosa , and Mycobacterium abscessus are common pathogens in the United States that can be found in drinking water. These organisms can persist despite disinfection, colonize plumbing, and proliferate under various temperature and flow conditions. Effective monitoring is crucial for public health protection and responding to outbreaks. Water monitoring methods typically include a concentration step, as the microbial load in drinking water is low. Centrifugation and filtration methods have traditionally been used in these workflows; however, these are very manual processes and are difficult to scale up in situations where high levels of testing are required. To address these limitations, we used Nanotrap® Microbiome Particles, magnetic hydrogel nanoparticles, previously validated in wastewater and clinical matrices(1), as an alternative concentration strategy for pathogen detection in tap water. Their use was assessed across multiple downstream analytical workflows, including dPCR, metagenomic sequencing, and culturing. POSTER SKU 44XXX Literature # WW-PO31451

The adoption of wastewater-based epidemiology (WBE) for monitoring SARS-CoV-2 has rapidly expanded worldwide, driven by strong evidence linking WBE and clinical case trends. The success of WBE for monitoring of SARS-CoV-2 prevalence in specific regions has led to a renewed interest in monitoring additional microorganisms, including yeast, such as Candida auris ( C. auris ). Transmission of Candida  species is oftentimes linked to physical contact with an infected host or through direct contact with a contaminated surface, particularly in hospitals and healthcare facilities. 1   C. auris  has also been observed to contain genes that confer antifungal resistance, 2  leading to the further need for broad surveillance of infectivity rates due to the limited treatment options available. Detecting these microbes also presents challenges; they contain strong cell structures and are difficult to lyse using common nucleic acid extraction workflows, and their concentration in wastewater is often very low due to infection control measures in healthcare settings. Culture-based methods have proven effective for detecting and characterizing C. auris in wastewater, improving the ability to study its infectivity at the community level. POSTER SKU 44XXX Literature # WW-PO31450

Drinking water treatment aims to stabilize bacterial communities in drinking water and to remove human pathogens from the system. Legionella pneumophila, Pseudomonas aeruginosa , and Mycobacterium abscessus are common pathogens in the United States that can be found in drinking water. These organisms can persist despite disinfection, colonize plumbing, and proliferate under various temperature and flow conditions. Effective monitoring is crucial for public health protection and responding to outbreaks. Water monitoring methods typically include a concentration step, as the microbial load in drinking water is low. Centrifugation and filtration methods have traditionally been used in these workflows; however, these are very manual processes and are difficult to scale up in situations where high levels of testing are required. To address these limitations, we utilized Nanotrap ® Microbiome Particles, magnetic hydrogel nanoparticles previously validated in wastewater and clinical matrices(1), as an alternative concentration strategy for pathogen detection in tap water. Their use was assessed across multiple downstream analytical workflows, including dPCR, metagenomic sequencing, and culturing. POSTER SKU 44XXX, 65XXX Lit # WW-PO31451

Sewer monitoring for antimicrobial resistance genes and organisms at healthcare facilities Rachel Poretsky, Dolores Sanchez Gonzalez, Adam Horton, Michael Schoeny, Chi-Yu Lin, Modou Lamin Jarju, Michael Secreto, Cecilia Chau, Ellen Gough, Erin Newcomer, Adit Chaudhary, Lisa Duffner, Nidhi Undevia, Angela Coulliette-Salmond, Amanda K. Lyons, Florence Whitehill, Mary K. Hayden, Stefan J. Green, Michael Y. Lin medRxiv 2025.03.16.25324079; doi: https://doi.org/10.1101/2025.03.16.25324079 This article is a preprint and has not been peer-reviewed [ what does this mean? ]. It reports new medical research that has yet to be evaluated and so should not be used to guide clinical practice. Abstract Surveillance of wastewater from healthcare facilities has the potential to identify the emergence of multidrug-resistant organisms (MDROs) of public health importance. Specifically, wastewater surveillance can provide sentinel surveillance of novel MDROs (e.g., emergence of Candida auris ) in healthcare facilities and could help direct targeted prevention efforts and monitor longitudinal effects. Several knowledge gaps need to be addressed before wastewater surveillance can be used routinely for MDRO surveillance, including determining optimal approaches to sampling, processing, and testing wastewater for MDROs. To this end, we evaluated multiple methods for wastewater collection (passive, composite, and grab), concentration (nanoparticles, filtration, and centrifugation), and PCR quantification (real-time quantitative PCR vs. digital PCR) for C. auris and 5 carbapenemase genes ( blaKPC, blaNDM, blaVIM, blaIMP , and blaOXA-48-like ) twice weekly for 6 months at a long-term acute care hospital in Chicago, IL. We also tested the effects of different transport and sample storage conditions on PCR quantification. All genes were detected in facility wastewater, with blaKPC being the most consistently abundant. Experiments were done in triplicate with gene copy, variance, and number of detections between triplicates used to determine method efficacy. We found that passive samples processed immediately using a combination of centrifugation followed by bead-beating and dPCR provided the most reliable results for detecting MDROs. We also present the trade-offs of different approaches and use culture and metagenomics to elucidate clinical relevance. This study establishes a practical approach for wastewater surveillance as a potential tool for public health monitoring of MDRO burden in healthcare facilities. Background Surveillance of wastewater from healthcare facilities has the potential to identify the emergence of multidrug-resistant organisms (MDROs) of public health importance [ 1 ]. Optimal approaches to sampling, processing, and testing wastewater need to be established before wastewater surveillance (WWS) can be used routinely for MDRO surveillance. Although genes of interest can be detected using similar molecular approaches in both wastewater samples and patient samples, there are multiple interrelated and unique challenges to WWS. First, wastewater is a complex medium, containing a mixture of organic matter, particles, microorganisms, and flow variability. Second, laboratory methods for WWS have not yet been standardized, leading to poor understanding of measurement uncertainties and the effects of wastewater composition on detection [ 2 – 4 ]. Finally, target abundance can hinder detection; previous metagenomics-based studies in municipal and hospital wastewater have shown that many antimicrobial resistance (AR) genes are present at low relative abundances [ 5 – 7 ]. Here, we present an evaluation of approaches to address these knowledge gaps, focusing on healthcare associated pathogens: Candida auris and carbapenemase-producing organisms (CPOs, represented by five target genes blaKPC, blaNDM, blaVIM, blaIMP , and blaOXA-48-like ). We collected wastewater from a region of the United States where C. auris and CPOs are endemic [ 8 – 11 ]. This study employed a longitudinal design at a single long-term acute care hospital over a period of 6 months. We compared three sample collection methods (passive, composite, and grab), three sample processing methods (magnetic nanoparticle [NP] concentration, InnovaPrep Concentrating Pipette [ICP] filtration, and centrifugation followed by bead-beating), and two different detection methods (quantitative real-time PCR [qPCR] and digital PCR [dPCR]) with MDRO diagnostic assays that have been developed and validated for use in clinical samples; therefore, additional evaluation is needed to optimize performance in wastewater samples. We report the findings of a series of seminal experiments that establish an effective protocol for surveillance of C. auris and CPOs in healthcare facility wastewater, helping to define the use of WWS as an important tool for monitoring MDRO burden in healthcare facilities. ARTICLE SKU 44XXX, 65XXX https://www.medrxiv.org/content/10.1101/2025.03.16.25324079v1

In this technical note, we compare the Nanotrap® Extracellular Vesicle A Particles to an ultrafiltration method for concentration of extracellular vesicles from the eluent of a qEV size exclusion column. TECH NOTE SKU # 552XX Lit. # PL-TN31418

In this technical note, we demonstrate improved qPCR detection of miR-16, a common microRNA present in extracellular vesicles, using magnetic Nanotrap® Extracellular Vesicle B Particles. TECH NOTE SKU # 552XX Lit. # PL-TN31419

In this study, we demonstrate that the Nanotrap® Protein Enrichment Affinity Kit (PEAK) is compatible with the PreOmics® iST kit for proteomic analysis of human plasma. TECH NOTE SKU # 34XXX Lit. # PL-TN31430

The Nanotrap® Protein Enrichment Affinity Kit (PEAK) uses magnetic hydrogel particle technology to capture and concentrate lowabundance proteins and peptides, leading to improved protein detection via LC-MS/MS. With three unique Nanotrap® Protein Particle chemistries available, researchers can easily adapt their approach based on specific protein targets or use case. Here we demonstrate that a simple, 45-minute enrichment process using the Nanotrap® PEAK is compatible with commercial digestion kits from four different vendors: Promega, Pierce, PreOmics, and Thermo Fisher Scientific. We also demonstrate that the Nanotrap PEAK significantly decreases the albumin levels in plasma samples. POSTER SKU # 34XXX Lit. # PL-PO31443

Detecting low-abundance biomarkers in complex biological samples is a persistent challenge in proteomics and clinical diagnostics. Conventional sample processing methods do not provide sufficient enrichment of biomarkers present in clinical sample matrices, which are often present at low concentrations. The Nanotrap® Protein Enrichment Affinity Kit (PEAK) enhances protein concentration and recovery. In this poster, we implemented the manual and automated Nanotrap® PEAK workflows, demonstrating a high-throughput method for protein biomarker enrichment that improves protein detection sensitivity from human K2EDTA plasma. POSTER SKU # 34XXX Lit. # PL-PO31444

Mass spectrometry-based proteomic analysis of plasma is a vital tool for biomarker discovery, yet it is hindered by high-abundance proteins, which can obscure the detection of low-abundance biomarkers. Nanotrap® Protein Enrichment Affinity Kit (PEAK)—simple, versatile, and easy-to-use kits that improve the detection of low-abundance proteins from multiple sample types. These kits utilize the Nanotrap® magnetic hydrogel particle technology to enrich low-abundance proteins from complex sample matrices. In this poster, three different Nanotrap® Protein Particle types and multiple particle combinations for plasma processing were evaluated. Each of the workflows offers unique benefits, allowing researchers to tailor their approach. Additionally, we investigated whether a simple 30-minute enrichment step using Nanotrap® PEAK would enhance protein identification in plasma, cerebrospinal fluid (CSF), and urine samples. POSTER SKU # 34XXX Lit. # PL-PO31442

In this study, we demonstrate that the Nanotrap® Protein Enrichment Affinity Kit (PEAK) is compatible with the Pierce™ In-Solution Tryptic Digestion and Guanidination Kit for proteomic analysis of human plasma. TECH NOTE SKU # 34XXX Lit. # PL-TN31431

Mpox (formerly known as monkeypox) has historically been present in western and central Africa. However, global outbreaks in May 2022 and August 2024 prompted the World Health Organization (WHO) to declare public health emergencies of international concern. The 2022 outbreak was linked to Clade II Mpox. The ongoing 2024 outbreak is driven by a newly identified strain of Clade I Mpox, known as Clade Ib. Clade I is believed to have a higher fatality rate than clade II. Clade Ib was first detected in South Kivu, in the Democratic Republic of the Congo, and now multiple strains of Clade Ib are circulating.(1,2)  Clade Ib Mpox is difficult to diagnose based on clinical symptoms alone, as its visual presentation is not distinct enough for reliable identification.(3) This variant includes a deletion in a genomic region previously targeted by diagnostic tests,(4) which may contribute to underreporting of cases. However, sequencing data from Clade Ib strains have enabled the development of a new PCR-based assay for detecting Mpox in wastewater. This assay targets a region of the genome that is not affected by the deletion, offering a promising tool for monitoring community infection levels and tracking the spread of the disease. POSTER SKU 44XXX