Detection Of Viruses In Foods Research Articles

Recent Advances in Biosensor Development for the Detection of Viral Particles in Foods: A Comprehensive Review.

Viral foodborne diseases cause serious harm to human health and the economy. Rapid, accurate, and convenient approaches for detecting foodborne viruses are crucial for preventing diseases. Biosensors integrating electrochemical and optical properties of nanomaterials have emerged as effective tools for the detection of viruses in foods. However, they still face several challenges, including substantial sample preparation and relatively poor sensitivity due to complex food matrices, which limit their field applications. Hence, the purpose of this review is to provide an overview of recent advances in biosensing techniques, including electrochemical, SERS-based, and colorimetric biosensors, for detecting viral particles in food samples, with emerging techniques for extraction/concentration of virus particles from food samples. Moreover, the principle, design, and advantages/disadvantages of each biosensing method are comprehensively described. This review covers the recent development of rapid and sensitive biosensors that can be used as new standards for monitoring food safety and food quality in the food industry.

Short Communication — Stability of “Free” Norovirus RNA on Fresh Produce

AbstractThe accuracy and dependability of results generated through molecular detection of specific target sequences of RNA, commonly used for detecting viruses in food, have been extensively debated within the scientific community. Such concerns have been raised by researchers, clients, and regulators alike, highlighting the need for further investigation and clarification. In particular, there has been debate about the possibility of molecular methods to detect enteric viruses, such as norovirus, in foods, producing false positive RT-PCR results due to the presence of “free” target RNA sequences in the food supply chain environment. This study aimed to investigate this issue by evaluating the recovery of “free” norovirus RNA from lettuce leaves. The “free” RNA was produced using heat treatment and chemical extraction methods. The study findings indicate that recovery of heat-extracted RNA from the lettuce leaves decreased markedly within 24 h, while recovery of chemically extracted RNA remained stable and even increased over time. The results of this study suggest that positive molecular detection of viruses in food is more likely to be associated with intact and potentially infectious virus particles, rather than “free” unprotected RNA. These findings have significant implications for the food industry and regulatory bodies in terms of the interpretation, accuracy and reliability of molecular detection methods for virus detection in food.

Inactivation of viruses related to foodborne infections using cold plasma technology

AbstractGlobally, there is a rise in day‐to‐day demand for minimally processed foods to supply nutritious, wholesomeness and safe foods to the consumers. Contamination of food by pathogens is a serious problem resulting in several outbreaks. Food pathogens like molds, bacteria were detectable and can be inactivated. The virus detection in foods is always a difficult task as their presence could not alter any noticeable change in the quality. Norovirus, Hepatitis A viruses are well‐known for their foodborne outbreaks and illnesses. Enveloped viruses are resistant and have the stability to the current traditional preservation methods due to the presence of a protective capsid layer and an envelope. The current thermal processing has shown significant effect on the product quality. The use of chemical disinfestation compounds is not suitable for food commodities. There is a need for alternative nonthermal food processing technologies for decontamination of food and food packages and preserving the food quality as well. Cold plasma is one of the emerging nonthermal, chemical‐free residues, and eco‐friendly technology widely being applied to the different food sectors. The main antiviral mechanism is the disruption of the capsid protein layer, the oxidation and denaturation of viral proteins. The method has also caused damage to the envelope layer and genetic material. This review focuses on cold plasma inactivation efficiency on different viruses.

Interlaboratory Evaluation of a Method for Quantification of Norovirus RNA as an Alternative Use for ISO 15216-1:2017 to Conduct Japan Baseline Survey of Oysters.

In this study, we aimed to investigate the standard method used for quantification of norovirus in oysters in Japan for the provisional adaptation of the method as an alternative to ISO 15216-1:2017, to conduct a Japan baseline survey of norovirus in oysters. For this purpose, the method provided by the Japan Committee for Standardization of Virus Detection in Food was subjected to an interlaboratory study to determine the performance characteristics of the standard method used in Japan. As a result, the theoretical limit of quantification for norovirus GI and GII in oysters by the standard method used in Japan was expected to be 1.92 and 1.85 log10 copies/g, respectively. The repeatability standard deviations (Sr) were 0.26 and 0.30 log10 copies/g for GI and GII, respectively, and the reproducibility standard deviations (SR) were 0.47 and 0.44 log10 copies/g for GI and GII, respectively. Through the interlaboratory study, we specified several critical points to obtain scientifically reliable results by using the standard method used in Japan. Especially, necessity for application of using process control virus was the most crucial point that needed to be improved. In addition, there are many participating laboratories that could not handle dilution of standard and quantify or detect the viruses in the test samples. To ensure scientifically reliable test result, capacity building of laboratories and implementation of proficiency testing should be considered for future tasks in combination with an application of process control materials in the method. On the assumption that the problems revealed in this study will be solved, the standard method used in Japan would be suitable for use in Japan baseline survey of norovirus in oysters, which will contribute to the international action against norovirus in oysters, led by the EU.

HAV detection from milk-based products containing soft fruits: Comparison between four different extraction methods

Virus detection in food requires appropriate elution and concentration techniques which need to be adapted for different food matrices. ISO/TS-15216-1:2017 and ISO/TS-15216-2:2019 describe standard methods for hepatitis A virus (HAV) research in some food only. Milk-based products containing one or more types of fruit are not covered by ISO procedures, even though they can be contaminated by fruit added to these products or by the food handlers. The aim of this work was to identify an efficient method for the detection of HAV in milk-based products. Four methods were tested to recover HAV from artificially contaminated milk, yoghurt and ice cream containing soft fruits. Results showed that the efficiency of the tested methods depends on the analyzed matrix. In milk we obtained a mean recovery from 13.4% to 1.9%; method based on high speed centrifuge gave the best values. The average recovery in yoghurt was between 3.3% and 114.4%, the latter value achieved by method with beef extract at 3% as eluent. Finally, two methods gave the best results in ice cream with similar recoveries: 29.1% and 27.7% respectively. The first method used glycine as eluent while the other one was based on high speed centrifugation. The ISO method has never proved to be the most efficient in the matrices studied. Therefore, based on the results obtained, a complete rethinking of the ISO method may be necessary to improve its recovery for some products such as milk, while only small changes would be sufficient for other products, such as yoghurt and ice cream.

Prevalence and evaluation strategies for viral contamination in food products: Risk to human health—a review

ABSTRACTNowadays, viruses of foodborne origin such as norovirus and hepatitis A are considered major causes of foodborne gastrointestinal illness with widespread distribution worldwide. A number of foodborne outbreaks associated with food products of animal and non-animal origins, which often involve multiple cases of variety of food streams, have been reported. Although several viruses, including rotavirus, adenovirus, astrovirus, parvovirus, and other enteroviruses, significantly contribute to incidence of gastrointestinal diseases, systematic information on the role of food in transmitting such viruses is limited. Most of the outbreak cases caused by infected food handlers were the source of 53% of total outbreaks. Therefore, prevention and hygiene measures to reduce the frequency of foodborne virus outbreaks should focus on food workers and production site of food products. Pivotal strategies, such as proper investigation, surveillance, and reports on foodborne viral illnesses, are needed in order to develop more accurate measures to detect the presence and pathogenesis of viral infection with detailed descriptions. Moreover, molecular epidemiology and surveillance of food samples may help analysis of public health hazards associated with exposure to foodborne viruses. In this present review, we discuss different aspects of foodborne viral contamination and its impact on human health. This review also aims to improve understanding of foodborne viral infections as major causes of human illness as well as provide descriptions of their control and prevention strategies and rapid detection by advanced molecular techniques. Further, a brief description of methods available for the detection of viruses in food and related matrices is provided.

Review: Approaches to the Viral Extraction, Detection, and Identification of Hepatitis Viruses, HAV and HEV, in Foods.

Although the incidence rate of hepatitis A virus (HAV) infection has been on the decline in developed countries, in part due to immunization availability, high profile outbreaks continue to be reported. Hepatitis E virus has been recognized as an emerging pathogen in industrialized countries. While associated with waterborne illnesses, particularly in undeveloped countries, several animal species have been identified as reservoirs for the virus. Consequently, the potential of zoonotic transmission exists as a function of the consumption of infected animals. In this review we provide a comparative overview of these two virus species with regard to their known virus properties, discuss extraction methodologies, and describe some basic principles and methodology applied toward the isolation of these viruses (as particles or their isolated genomes) from food commodities. We also discuss the challenges that remain as experimental hurdles to extraction of such viruses from food. As HAV has been the most extensively studied with regard to virus detection in foods, it often serves as a model virus for current and future development of sample preparation methodology for foodborne virus detection. Lastly, we discuss the application and role of current and developing technologies in the post-extraction detection and identification of these viruses from foods.

Analytical methods for the detection of viruses in food by example of CCL-3 bioagents

This critical review presents challenges and strategies in the detection of viral contaminants in food products. Adenovirus, caliciviruses, enteroviruses, and hepatitis A are emerging contaminant viruses. These viruses contaminate a variety of food products, including fruits, vegetables, shellfish, and ready-to-eat processed foods. The diversity of targets and sample matrices presents unique challenges to virus monitoring that have been addressed by a wide array of processing and detection methods. This review covers sample acquisition and handling, virus recovery/concentration, and the determination of targets using molecular biology and mass-spectrometric approaches. The concentration methods discussed include precipitation, antibody-based concentration, and filtration; the detection methods discussed include microscopy, polymerase chain reaction, nucleic acid sequence-based amplification, and mass spectrometry.

食品のウイルス検査の現状と課題

食品のウイルス検査の現状と課題

Achievement V – Methods for breaking the transmission of pathogens along the food chain: Detection of viruses in food

Traditionally the focus for control of food-borne disease has been bacteria. During the last decade viruses have emerged as important sources of food borne human disease. Since the traditional bacteriological indicators, are not reliable for viral contamination, new methods are needed. PCR has enhanced the detection of virus in food. A challenge for developing detection reliable methods for viruses in food is that food matrices vary in composition, high sequence variability and inhibitors may be present. Therefore it is necessary to develop assays that have high diagnostic sensitivity, are broad and robust, and combine sample concentration and removal of inhibitors.

Nucleic Acid Amplification-Based Methods for Detection of Enteric Viruses: Definition of Controls and Interpretation of Results

The most effective methods for virus detection in food and environmental samples are those based on nucleic acid amplification. Complex methods must be applied by the analyst in order to control for false negative results of virus detection assays in those samples that may be contaminated by virus concentrations above the detection level. The verification of analytical results is a necessity and this depends on using an appropriate suite of controls to monitor the efficacy of the critical steps in the method and allow correct result interpretations to be made. We describe the suite of controls necessary for analysing food and environmental samples for the presence of enteric viruses by nucleic acid amplification-based methods. To exclude false negative and positive interpretations of results, the inclusion of this suite of controls will be essential when considering incorporating monitoring of viruses in food or environmental safety management plans.

Analytical Methods for Virus Detection in Water and Food

Potential ways to address the issues that relate to the techniques for analyzing food and environmental samples for the presence of enteric viruses are discussed. It is not the authors’ remit to produce or recommend standard or reference methods but to address specific issues in the analytical procedures. Foods of primary importance are bivalve molluscs, particularly, oysters, clams, and mussels; salad crops such as lettuce, green onions and other greens; and soft fruits such as raspberries and strawberries. All types of water, not only drinking water but also recreational water (fresh, marine, and swimming pool), river water (irrigation water), raw and treated sewage are potential vehicles for virus transmission. Well over 100 different enteric viruses could be food or water contaminants; however, with few exceptions, most well-characterized foodborne or waterborne viral outbreaks are restricted to hepatitis A virus (HAV) and calicivirus, essentially norovirus (NoV). Target viruses for analytical methods include, in addition to NoV and HAV, hepatitis E virus (HEV), enteroviruses (e.g., poliovirus), adenovirus, rotavirus, astrovirus, and any other relevant virus likely to be transmitted by food or water. A survey of the currently available methods for detection of viruses in food and environmental matrices was conducted, gathering information on protocols for extraction of viruses from various matrices and on the various specific detection techniques for each virus type.

Food-Borne Viruses: Progress and Challenges

One question that is well known to persons working with food virology, and especially noroviruses, is “How important are these viruses, actually?” Frequently you feel somewhat uneasy when you start to reply, talking about the “trivial illness” of gastroenteritis, continuing on to the economic impact caused by the huge number of cases, and finally ending up admitting that you really do not know the answer. This book does not provide an answer to this difficult question either, but it does give you, among other things, an introduction to the pitfalls in trying to “estimate the burden of an underreported disease.” One conclusion is that we probably underestimate its importance. The book consists of 10 chapters written by persons well known to food virologists as experts in their fields. It is well organized, well written, and presents the history of foodborne viruses, the state of the art, and a glance into the future. The history of food virology is given by Dean O. Cliver, one of the pioneers in this area, who spices up his contribution by sharing interesting personal experiences with the reader. Most of the focus in the book is, naturally, directed toward norovirus and hepatitis A virus. These viruses are covered by up-to-date presentations on molecular biology, clinical disease and diagnostics, pathogenesis and immunity, epidemiology, and detection in food matrixes. Recommendations are also given about which foods should be tested and under which circumstances. Although protocols are not presented, many references are listed. The molecular revolution has improved the detection of viruses that are impossible or difficult to propagate in cell culture, such as norovirus and hepatitis A virus, and reverse transcription–PCR has become the standard method for virus detection in food. However, sensitive detection methods are still lacking. This is one of the recurrent topics in the book because it affects outbreak investigation, risk assessment, and risk management. The need for standardization for virus detection in food is stated, as is the proper use of controls. Nobody will argue against detection of false-negative and false-positive results, but some thoughts on the cost-benefit aspects regarding accurate quantification of virus genomes in foods would have been interesting. The book provides a broad overview of the present situation and gives a good introduction to the topic. Factors that may contribute to the emergence of new viral diseases are also discussed, including demographic and pathogen-related changes. This is interesting reading because virus families and viral evolution are discussed within the context of foodborne transmission. The editors encourage the reader to enjoy the book, and, most of the time, I did.

Food-borne viruses: progress and challenges

Table of Contents 1. Historic Overview of Food Virology, Dean O. Cliver 2. Food-Borne Viruses-State of the Art, Marc-Alain Widdowson and Jan Vinje 3. Enterically Transmitted Hepatitis, Rakesh Aggarwal 4. The Challenge of Estimating the Burden of an Underreported Disease, Sarah J. O'Brien 5. Emerging Food-Borne Viral Diseases, Erwin Duizer and Marion Koopmans 6. Viral Evolution and Its Relevance for Food-Borne Virus Epidemiology, Esteban Domingo and Harry Vennema 7. Rethinking Virus Detection in Food, Rosa M. Pinto and Albert Bosch 8. Binding and Inactivation of Viruses on and in Food, with a Focus on the Role of the Matrix, Francoise S. Le Guyader and Robert L. Atmar 9. Codex' Risk Analysis Framework To Reduce Virus in Food Risks, Jaap Jansen 10. Risk Assessment of Viruses in Food: Opportunities and Challenges, Arie H. Havelaar and Saskia A. Rutjes

Viruses transmitted through the food chain: a review of the latest developments.

Abstract The emergence of severe acute respiratory syndrome (SARS), Nipah virus and H5N1 has sparked fears of food-borne dissemination of viruses, resulting in an increased focus on microbiological food safety. Norovirus (NoV) and hepatitis A virus (HAV) have been recognized as priorities based on their illness burden, and as models for prevention studies. In addition, hepatitis E virus is considered an emerging food-borne pathogen. Systematic surveillance for food-borne viral diseases, where present, has to deal with underreporting, with available data biased towards developed countries, making true estimates of the burden of illness challenging. Nevertheless, food-borne viral outbreaks are often reported, whereas current measures undertaken by food safety management organizations are focused primarily on bacterial contamination and are insufficient for viruses in food. There has been a shift towards molecular techniques for the diagnosis and detection of viruses in food, but a definite link to food is still hard to establish owing to the low levels of viruses in foods. Moreover, multiple strains may be involved in sewage-contaminated foods, with the risk of recombination and development of novel strains after ingestion. To enable monitoring of food and food safety, sensitive routine methods are needed, and methods for detection of NoV and HAV in risk foods (shellfish, soft fruit and leafy greens) are currently validated. Norovirus cannot be cultured, but the murine NoV may be a surrogate for studying NoV survival. In all, significant progress has been made in understanding food-borne viral infections, but clear data gaps still exist and should be the focus of future research.

Hepatitis A virus detection in food: current and future prospects

Hepatitis A virus (HAV) is responsible for around half of the total number of hepatitis infections diagnosed worldwide. HAV infection is mainly propagated via the faecal-oral route and as a consequence of globalisation, transnational outbreaks of foodborne infections are reported with increasing frequency. Molecular procedures are now available and should be employed for the direct surveillance of HAV in food and environmental samples.

Internationally Distributed Frozen Oyster Meat Causing Multiple Outbreaks of Norovirus Infection in Australia

Between November 2003 and January 2004, outbreaks of norovirus in 3 Australian jurisdictions involving 83 cases of illness were associated with imported oyster meat. Cohort studies were conducted in 2 jurisdictions to identify relative risks of illness for the consumption of oysters. A case series was conducted in the third jurisdiction. The cohort studies conducted in the first 2 jurisdictions identified relative risks of illness of 17 (95% confidence interval, 5-51) and 35 (95% confidence interval, 5-243), respectively, for the consumption of oysters. Multiple strains of norovirus were detected in fecal specimens from 8 of 14 patients and in 1 of the 3 batches of implicated oyster meat using seminested reverse-transcriptase polymerase chain reaction methods. Traceback investigations revealed that all oyster meat was harvested from the same estuary system in Japan within the same month. These outbreaks demonstrate the potential of foodborne disease to spread internationally and the need for national and international collaboration to investigate such outbreaks. Foodborne illness related to norovirus is underestimated because of underreporting of human cases and challenges in laboratory detection of viruses in foods, both of which can delay public health action.

Detection of Noroviruses in Foods: A Study on Virus Extraction Procedures in Foods Implicated in Outbreaks of Human Gastroenteritis

Disease outbreaks in which foods are epidemiologically implicated as the common source are frequently reported. Noroviruses and enteric hepatitis A viruses are among the most prevalent causative agents of foodborne diseases. However, the detection of these viruses in foods other than shellfish is often time-consuming and unsuccessful. In this study, three virus concentration methods were compared: polyethylene glycol (PEG) plus NaCl, ultracentrifugation, and ultrafiltration. Two RNA extraction methods, TRIzol and RNeasy Mini Kit (Qiagen), were compared for detection of viruses in whipped cream and lettuce (as representatives of the dairy and vegetable-fruit food groups, respectively). A seeding experiment with canine calicivirus was conducted to determine the efficiency of each virus extraction procedure. The PEG-NaCl-TRIzol method was most efficient for the detection of viruses in whipped cream and the ultracentrifugation–RNeasy–Mini Kit procedure was best for detection on lettuce. Based on the seeding experiments, food items implicated in norovirus-associated gastroenteritis outbreaks were subjected to the optimal procedure for a specific composition and matrix. No noroviruses were detected in the implicated food items, possibly because the concentration of virus on the food item was too low or because of the presence of inhibitory factors. For each food group, a specific procedure is optimal. Inhibitory factors should be controlled in these procedures because they influence virus detection in food.

A Polymerase Chain Reaction–Based Method for the Detection of Hepatitis A Virus in Produce and Shellfish

Outbreaks of gastroenteritis that are suspected to be of viral origin are on the rise. Thus, there is a need for regulatory agencies entrusted with food safety to develop adequate techniques for the detection of viruses in foods. We have established a general procedure for the detection of hepatitis A virus (HAV) in shellfish that, with minor modifications, is also applicable to fresh produce such as cilantro. Total RNA was isolated from shellfish or cilantro, followed by isolation of poly(A)-containing RNA. Because HAV genomic RNA contains a poly(A) tail, the isolation of poly(A)-containing RNA also enriches HAV genomic RNA. Reverse transcription was used to convert the RNA to cDNA, and then amplification was carried out by polymerase chain reaction (PCR). Reamplification with internal primers was used to improve the quality and the quantity of amplified DNA, allowing for post-PCR analysis such as sequence identification of the viral strain. With this procedure, multiple samples could be analyzed in four working days by a single trained individual. The nominal sensitivity of detection of the procedure was 0.15 TCID50 (50% tissue culture infective dose) per 0.62g of tissue with a test virus. The direct RNA isolation protocol avoided pitfalls associated with whole-virus purification procedures by replacing virus precipitation steps involving polyethylene glycol and Procipitate with phenol extraction. The method is straightforward and reliable. We successfully used this procedure to detect naturally occurring HAV in clams involved in a gastroenteritis outbreak, as well as in cilantro artificially contaminated with a test virus.