Absolute quantification of norovirus capsid protein in food, water, and soil using synthetic peptides with electrospray and MALDI mass spectrometry

Abstract

Norovirus infections are one of the most prominent public health problems of microbial origin in the U.S. and other industrialized countries. Surveillance is necessary to prevent secondary infection, confirm successful cleanup after outbreaks, and track the causative agent. Quantitative mass spectrometry, based on absolute quantitation with stable-isotope labeled peptides, is a promising tool for norovirus monitoring because of its speed, sensitivity, and robustness in the face of environmental inhibitors. In the current study, we present two new methods for the detection of the norovirus genogroup I capsid protein using electrospray and matrixassisted laser desorption/ionization (MALDI) mass spectrometry. The peptide TLDPIEVPLEDVR was used to quantify norovirus-like particles down to 500 attomoles with electrospray and 100 attomoles with MALDI. With MALDI, we also demonstrate a detection limit of 1 femtomole and a quantitative dynamic range of 5 orders of magnitude in the presence of an environmental matrix effect. Due to the rapid processing time and applicability to a wide range of environmental sample types (bacterial lysate, produce, milk, soil, and groundwater), mass spectrometry-based absolute quantitation has a strong potential for use in public health and environmental sciences.

Evaluation of Virucidal Efficacy of Human Norovirus Using Combined Sprayed Slightly Acidic Electrolyzed Water and Ultraviolet C-Light-Emitting Diode Irradiation Treatment Based on Optimized Capture Assay for Quantitative RT-qPCR

“…HuNoV GII.4 is recognized as a major cause of acute gastroenteritis outbreaks, and sporadic illnesses caused by HuNoV GII.4 are reported ( Hartmann et al, 2015 ). To eliminate HuNoV contamination, a potentially contaminated matrix should be disinfected properly using various methods.…”

“…Human norovirus (HuNoV), which has characteristics such as high viral load excretion, tenacious environmental sustainability, rapid propagation, and short-term immunity, causes sporadic cases of viral gastroenteritis. Owing to its high contagiousness and rapid spread around the world, it infects humans regardless of gender or age ( Hartmann et al, 2015 ). HuNoV infection is the most common cause of acute gastroenteritis, causing approximately 680 million cases per year ( CDC, 2021 ).…”

Mass Spectrometry Approaches for SARS-CoV-2 Detection: Harnessing for Application in Food and Environmental Samples

“…The most widely used mass spectrometric approach for pathogen identification is matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) [ 16 , 17 , 18 , 19 ]. The success of this platform is due to the multiple in vitro protocols developed for bacteria biotyping methods, many of which have been approved by regulatory agencies such as the US Food and Drug Administration (FDA).…”

Optimizing Operating Parameters of Electric Ultra-Low Volume Sprayer with Slightly Acidic Electrolyzed Solution for Efficient Virucidal Activity on Environmental Surfaces

“…Human noroviruses (Hu-NoVs), characterized by high infectivity among humans, short-term immunity, tenacious environmental persistence, and high viral load excretion, cause sporadic cases of nonbacterial gastroenteritis and represent the most significant etiologic agent of human epidemics, regardless of age or sex, and likely owing to their high-transmissibility across industrialized nations [ 1 ]. Moreover, the Centers for Disease Control and Prevention (CDC) reported that, annually, Hu-NoV infections cause an estimated 200,000 deaths, with more than 590,000,000 cases worldwide [ 2 ].…”