Tuberculosis (TB) remains a leading global health concern, causing approximately 1.4 million deaths annually.1 Early and accurate detection of Mycobacterium tuberculosis in complex clinical samples, such as sputum, is critical for effective diagnosis and treatment. Traditional culture methods, while highly specific, are slow and labor-intensive. Molecular approaches such as qPCR and sequencing provide faster results but are often hindered by inefficient mycobacterial recovery and the presence of inhibitors in sputum.(2) To address these limitations, we investigate the use of the Nanotrap® Microbiome B Particles. The particles are composed of a hydrogel matrix and functionalized with affinity bait molecules that capture and concentrate intact microbes from various sample matrices. We demonstrate that this approach can effectively be used to capture and concentrate Mycobacterium  directly from sputum, improving sensitivity and reducing workflow complexity. POSTER SKU 65XXX

Wastewater-based epidemiology (WBE) has emerged as a crucial tool for monitoring infectious diseases at the population level, providing early indicators of viral transmission and evolution. Influenza A viruses, known for their seasonal epidemics and pandemic potential, pose unique challenges for wastewater surveillance due to their low abundance and susceptibility to environmental degradation (1, 2). Traditional sequencing approaches for influenza A in wastewater often yield incomplete genomes due to low viral titers and matrix inhibitors, limiting their utility for public health surveillance(3).  This study presents an enhanced workflow combining Nanotrap® Microbiome A Particles for viral enrichment with the NEBNext iiMS Influenza A DNA Library Prep protocol, leveraging Oxford Nanopore sequencing for rapid whole-genome characterization. By integrating targeted cDNA synthesis and native barcoding, this approach enables same-day sequencing with improved genome coverage, offering a scalable and high-throughput solution for monitoring influenza A in wastewater. POSTER SKU 44XXX

Wastewater-based epidemiology (WBE) has become a vital tool for tracking SARS-CoV-2 outbreaks, which has led to interest in expanded pathogen tracking. Extensive research has been conducted on the persistence of human enteric viruses transmitted via the fecal-oral route in wastewater. Common targets for multi-pathogen environmental surveillance have been identified as enteric viruses, Salmonella enterica serovar Typhi, Vibrio cholerae , SARS-CoV-2, hepatitis A and E virus, and measles virus. A report from the Centers for Disease Control and Prevention (CDC) indicated 58 confirmed cases of measles in the United States in early 20244, leading to an increased need for community-based detection. Nanotrap® Microbiome Particles capture and concentrate a variety of pathogens prior to nucleic extraction and detection. Using standard Nanotrap Particle workflows, we aim to demonstrate the utility of this method to capture and concentrate Measles morbillivirus , Hepatitis A Virus (HAV), Enterovirus 71 (EV71), and Salmonella enterica from wastewater samples as a tool for bio-surveillance applications and to facilitate rapid response for potential infectious disease outbreaks.

A clinician outreach and communication report from the Centers for Disease Control and Prevention (CDC) indicated 58 confirmed cases of measles in the United States from January 1, 2024 to March 14, 2024¹. Community surveillance of measles plays a crucial role in tracking and preventing the spread of measles to unvaccinated individuals. This study aimed to determine whether the measles virus could be captured and detected in wastewater influent samples using Nanotrap® Microbiome A Particles and Nanotrap® Enhancement Reagent 3. TECHNICAL NOTE SKU 10XXX SKU 55XXX

This study evaluated the application of Nanotrap® Microbiome Particles to improve the recovery and culturing of Legionella pneumophila from tap water samples. Traditional filtration-based methods often struggle with low bacterial concentrations or require large sample volumes, limiting their utility in environmental monitoring. By spiking 35 mL tap water samples with varying concentrations of L. pneumophila , this study demonstrated that Nanotrap Microbiome Particles enabled the detection of viable bacteria at 0.1 cells/mL. In comparison, a CDC filtration method using 35 mL tap water and a 0.22 μm polycarbonate filter achieved growth only at concentrations ≥1 cell/mL. Colony Growth on BCYE Agar Plates Depicting the Results of Legionella pneumophila Detection TECHNICAL NOTE SKU 44XXX Lit. # TW-TN31414

Monitoring antimicrobial resistance (AMR) genes in agricultural environments is important for understanding and curbing the spread of resistance within ecosystems. Livestock waste, such as fecal slurry from pigs and cattle manure, often harbors AMR genes, which can be transferred to pathogenic bacteria, posing risks to both animal and human health. (1-3) Agricultural waste can also contain zoonotic pathogens like viruses and parasites, further compounding health risks. The widespread use of antibiotics in livestock contributes to the presence of resistance genes, such as tetM (tetracycline resistance) and sul2 (sulfonamide resistance), in livestock waste samples. These genes can spread to other bacteria through horizontal gene transfer, exacerbating the AMR threat. (4) Manure application on agricultural fields disseminates these genes into the soil microbiome, potentially impacting both animal and human health. (2,4) In addition to AMR genes, zoonotic pathogens in livestock waste, including viruses and parasites like Cryptosporidium and Giardia, pose significant health risks. Viral infections can lead to disease outbreaks in livestock, affecting both productivity and increasing the risk of zoonotic transmission to humans, especially those in close contact with animals. Parasites like Giardia also present direct risks to human health through contaminated water or handling infected animals. Effective detection and monitoring of these pathogens are important in preventing outbreaks and managing zoonotic transmission risks. The combined threat of AMR genes, zoonotic viruses, and parasites in livestock waste highlights the need for broad surveillance programs. These programs can guide interventions to reduce the risks associated with using livestock waste in agriculture, particularly within the One Health framework, which emphasizes the interconnectedness of human, animal, and environmental health. (1-3) Nanotrap Microbiome Particles enhance the capture and concentration of antimicrobial resistance (AMR) genes from livestock samples. APPLICATION NOTE SKU 44XXX, 65XXX, 10XXX Lit # SL-AN31391

Key Advantages Nanotrap Particles enable simple, automated methods for bio-surveillance of a broad spectrum of infectious diseases, including hepatitis A. The automated Nanotrap Particle method is compatible with several large-volume sample types, including produced wash water and wastewater. The automated Nanotrap Particle method offers equivalent or better hepatitis A viral detection from wastewater samples, as compared to an HA filtration method. Introduction Hepatitis A is an acute inflammatory liver disease which results from infection by the hepatitis A virus (HAV). It is faeco-orally spread, and its severity is age dependent. Recently, cases of acute hepatitis of unknown origin among young children have been reported from 12 countries around the world. Routine food safety testing can be used to effectively identify HAV present on foods, while wastewater surveillance for HAV can provide an indication of infection levels and affected geographical locations. Municipal wastewater harbors a variety of pathogenic viruses. Extensive research has been conducted on the persistence of human enteric viruses, which are transmitted via the fecal-oral route, in wastewater and in the aquatic environment. Wastewater surveillance is appealing because a single sample can ‘test’ a large population, it can enable detection from both asymptomatic and symptomatic individuals, and it can provide an indication of changing trends in population infection levels about 7 days ahead of clinical testing. CDC and HHS, in collaboration with agencies throughout the federal government, implemented the National Wastewater Surveillance System (NWSS) in response to the COVID-19 pandemic as a resource to help public health officials and communities utilize wastewater data to guide community level response and decision making. There is an interest in using the infrastructure in place to detect and monitor other common and potentially deadly pathogens. Nanotrap Particles have demonstrated sensitive, rapid, and easy-to-use automated and manual methods for wastewater SARS-CoV-2 testing. In this application note, we show that Nanotrap® Microbiome A Particles can also capture HAV from wastewater samples and from simulated agricultural produce wash samples. Nanotrap Microbiome A Particles outperformed the HA Filtration method for HAV capture and concentration in two spiked wastewater samples. APPLICATION NOTE SKU 10XXX SKU 44XXX

Mass-spectrometry-based proteomic analysis of biological samples is a powerful tool for the identification of potential biomarkers. Cerebrospinal fluid (CSF) is a common sample type used in proteomic analyses because of its clinical relevance and low protein complexity. However, high abundances of albumin and immunoglobin proteins can mask the detection of lower abundance proteins. (1) In this application note, we demonstrate how to use the Nanotrap® Protein Enrichment Affinity Kits (PEAK) to manually process CSF samples. We compare the number of unique protein identifications obtained from a CSF sample using different Nanotrap Protein Particle workflows to the same CSF sample processed without using the Nanotrap PEAK. We also compare the overlap of unique protein identifications for these different workflows. The Venn diagram illustrates the overlap of unique protein identifications (protein IDs) for the 1-particle Method using each of the three different Nanotrap Protein Particle types. APPLICATION NOTE SKU # 34XXX Lit. # CSF-AN31394

Droplet digital PCR is an attractive choice for rare mutation detection during liquid biopsy studies due to its powerful sensitivity, ease of use and data analysis, and generally lower price point than other common detection technologies such as next-generation sequencing. In this study, the NEAT Liquid Biopsy Kit was compared to a competitor kit using Bio-Rad ddPCR™ KRAS G12/G13 Screening Kit to detect KRAS mutations in plasma spiked with liquid biopsy standards at varying allele frequency. Figure 1. Number of Droplets Positive for KRAS G12/G13 Mutations from 4 mL Spiked Plasma TECHNICAL NOTE SKU 77XXX

In this application note, we use Sypro-Ruby staining of protein gels to demonstrate that each of the three Nanotrap Protein Particle types captures and concentrates proteins from K2EDTA plasma samples and reduces the presence of the highly abundant albumin protein. Sypro-Ruby staining images demonstrate that Nanotrap Protein Particles A, B, and C capture proteins across different size ranges and reduce the abundance of high molecular weight proteins. APPLICATION NOTE SKU # 34XXX

In this application note, we investigate the compatibility of Nanotrap® Particles with wastewater samples treated with Wastewater Stabilization Buffer. Additionally, we studied the claimed seven-day stability at room temperature for wastewater samples treated with Wastewater Stabilization Buffer using Nanotrap Particles for viral and bacterial target concentration. Confirming the stability supports the development of robust and efficient workflows for environmental monitoring and public health surveillance, particularly in the context of detecting and analyzing pathogens in wastewater. Demonstrating compatibility of Zymo Research’s Wastewater Stabilization Buffer with Nanotrap Particles for both viral and bacterial targets. APPLICATION NOTE SKU 44XXX, 65XXX

Over 200,000 new lung cancer patients are diagnosed every year, with over 150,000 deaths from the disease, making it one of the most lethal cancers. (1) Non-small cell lung cancer (NSCLC) makes up 85% of lung cancer patients in the US, with lung adenocarcinomas being the most common histological subtype of NSCLC. Lung adenocarcinomas are frequently characterized by KRAS mutations, which appear to correlate with disease aggressiveness. KRAS is also useful for predicting therapeutic response.(2) KRAS mutations can arise at any point during cancer progression, and detection of mutations as they appear is crucial to understanding the disease. Liquid biopsy of patient plasma is often used to understand disease progression in NSCLC patients, as mutations present in solid tumors can often be detected in plasma samples in low amounts. This non-invasive approach allows for more frequent sampling than traditional tissue biopsies but requires the detection of very low-frequency alleles. In this study, plasma from six NSCLC patients with known KRAS mutations was extracted by two different methods and the digital PCR (dPCR) results were compared. Quantification of wild-type KRAS in clinical plasma samples. APPLICATION NOTE SKU 77XXX