scholarly journals Evaluation of a New Environmental Sampling Protocol for Detection of Human Norovirus on Inanimate Surfaces

2015 ◽  
Vol 81 (17) ◽  
pp. 5987-5992 ◽  
Author(s):  
Geun Woo Park ◽  
David Lee ◽  
Aimee Treffiletti ◽  
Mario Hrsak ◽  
Jill Shugart ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Clinical Samples ◽  
Environmental Sampling ◽  
Viral Gastroenteritis ◽  
Human Norovirus ◽  
Drying Time ◽  
Viral Loads ◽  
Toilet Seat ◽  
Outbreak Investigations ◽  
Norovirus Gii

ABSTRACTInanimate surfaces are regarded as key vehicles for the spread of human norovirus during outbreaks. ISO method 15216 involves the use of cotton swabs for environmental sampling from food surfaces and fomites for the detection of norovirus genogroup I (GI) and GII. We evaluated the effects of the virus drying time (1, 8, 24, or 48 h), swab material (cotton, polyester, rayon, macrofoam, or an antistatic wipe), surface (stainless steel or a toilet seat), and area of the swabbed surface (25.8 cm2to 645.0 cm2) on the recovery of human norovirus. Macrofoam swabs produced the highest rate of recovery of norovirus from surfaces as large as 645 cm2. The rates of recovery ranged from 2.2 to 36.0% for virus seeded on stainless-steel coupons (645.0 cm2) to 1.2 to 33.6% for toilet seat surfaces (700 cm2), with detection limits of 3.5 log10and 4.0 log10RNA copies. We used macrofoam swabs to collect environmental samples from several case cabins and common areas of a cruise ship where passengers had reported viral gastroenteritis symptoms. Seventeen (18.5%) of 92 samples tested positive for norovirus GII, and 4 samples could be sequenced and had identical GII.1 sequences. The viral loads of the swab samples from the cabins of the sick passengers ranged from 80 to 31,217 RNA copies, compared with 16 to 113 RNA copies for swab samples from public spaces. In conclusion, our swab protocol for norovirus may be a useful tool for outbreak investigations when no clinical samples are available to confirm the etiology.

scholarly journals Efficacy of Neutral Electrolyzed Water for Inactivation of Human Norovirus

2017 ◽  
Vol 83 (16) ◽  
Author(s):  
Eric Moorman ◽  
Naim Montazeri ◽  
Lee-Ann Jaykus
Keyword(s):  
Stainless Steel ◽  
Receptor Binding ◽  
Electrochemical Activation ◽  
Viral Gastroenteritis ◽  
Human Norovirus ◽  
Electrolyzed Water ◽  
Astm Method ◽  
Steel Surfaces ◽  
Soil Load ◽  
The Impact

ABSTRACT Human norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Persistence on surfaces and resistance to many conventional disinfectants contribute to widespread transmission of norovirus. We examined the efficacy of neutral electrolyzed water (NEW; pH 7) for inactivation of human NoV GII.4 Sydney in suspension (ASTM method 1052-11) and on stainless steel surfaces (ASTM method 1053-11) with and without an additional soil load. The impact of the disinfectant on viral capsid was assessed using reverse transcriptase quantitative PCR (RT-qPCR; with an RNase pretreatment), SDS-PAGE, transmission electron microscopy, and a histo-blood group antigen (HBGA) receptor-binding assay. These studies were done in parallel with those using Tulane virus (TuV), a cultivable human NoV surrogate. Neutral electrolyzed water at 250 ppm free available chlorine produced a 4.8- and 0.4-log10 reduction in NoV genome copy number after 1 min in suspension and on stainless steel, respectively. Increasing the contact time on surfaces to 5, 10, 15, and 30 min reduced human NoV genomic copies by 0.5, 1.6, 2.4, and 5.0 log10 and TuV infectious titers by 2.4, 3.0, 3.8, and 4.1 log10 PFU, respectively. Increased soil load effectively eliminated antiviral efficacy regardless of testing method and virus. Exposure to NEW induced a near complete loss of receptor binding (5 ppm, 30 s), degradation of VP1 major capsid protein (250 ppm, 5 min), and increased virus particle aggregation (150 ppm, 30 min). Neutral electrolyzed water at 250 ppm shows promise as an antinoroviral disinfectant when used on precleaned stainless steel surfaces. IMPORTANCE Norovirus is the leading cause of acute viral gastroenteritis worldwide. Transmission occurs by fecal-oral or vomitus-oral routes. The persistence of norovirus on contaminated environmental surfaces exacerbates its spread, as does its resistance to many conventional disinfectants. The purpose of this research was to evaluate the antinoroviral efficacy of neutral electrolyzed water (NEW), a novel chlorine-based disinfectant that can be used at reduced concentrations, making it more environmentally friendly and less corrosive than bleach. An industrial-scale electrochemical activation device capable of producing relatively stable electrolyzed water at a wide pH range was used in this study. Experiments showed that 250 ppm NEW effectively eliminated (defined as a 5-log10 reduction) human norovirus GII.4 Sydney (epidemic strain) on clean stainless steel surfaces after a 30-min exposure. Supporting studies showed that, like bleach, NEW causes inactivation by disrupting the virus capsid. This product shows promise as a bleach alternative with antinoroviral efficacy.

scholarly journals Development of a Recombinase-Aided Amplification Assay for Rapid Detection of Human Norovirus GII.4

2020 ◽  
Author(s):  
Zhiwei Qin ◽  
Liang Xue ◽  
Weicheng Cai ◽  
Junshan Gao ◽  
Yueting Jiang ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Enzyme Reaction ◽  
Health Concern ◽  
Clinical Samples ◽  
Nucleic Acid Detection ◽  
Analytical Sensitivity ◽  
Human Norovirus ◽  
Amplification Assay ◽  
Stool Samples ◽  
Norovirus Gii

Abstract Background: Human noroviruses are one of the main causes of foodborne illnesses and represent a serious public health concern. Rapid and sensitive assays for human norovirus detection are undoubtedly necessary for clinical diagnosis, especially in regions without more sophisticated equipment.Method: The recombinase-aided amplification (RT-RAA) is a fast, robust and isothermal nucleic acid detection method based on enzyme reaction. This method can complete the sample detection at 39°C in 30 minutes. In this study, we successfully established a rapid reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of human norovirus GII.4 and applied this assay to clinical samples, as well as comparison with commercial RT-qPCR.Results: An analytical sensitivity detection of the RT-RAA at a 95% probability of 3.425 log10 genomic copies (LGC)/reaction. Moreover, no cross-reaction was observed with other norovirus genogroups and other common foodborne viruses. Stool samples were examined by RT-RAA and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Compared with RT-qPCR, kappa values for human norovirus detection with RT-RAA were 0.894 (p < 0.001), indicating that both assays were in agreement.Conclusion: This RT-RAA assay provides a rapid, specific, and sensitive assay for human norovirus detection and is suitable for clinical testing.

Real-time molecular beacon NASBA for rapid and sensitive detection of norovirus GII in clinical samples

2009 ◽  
Vol 55 (12) ◽  
pp. 1375-1380 ◽  
Author(s):  
Safaa Lamhoujeb ◽  
Hugues Charest ◽  
Ismail Fliss ◽  
Solange Ngazoa ◽  
Julie Jean
Keyword(s):  
Real Time ◽  
Molecular Beacon ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Reading Frame ◽  
Viral Loads ◽  
Reverse Transcription Pcr ◽  
Highly Sensitive ◽  
Nasba Assay ◽  
Norovirus Gii

To improve the sensitivity and efficiency of the real-time nucleic acid sequence based amplification (NASBA) assay targeting the open reading frame 1–2 (ORF1–ORF2) junction of the norovirus (NoV) genome, a selection of clinical samples were analyzed. The assay results were compared with those of TaqMan and conventional reverse transcription PCR (RT-PCR) and a commercial enzyme-linked immunoassay (ELISA) for the specific detection of GII NoV in 96 fecal samples. Based on end-point dilution, the two real-time assays had similar sensitivities (0.01 particle detectable units), two log10cycles greater than that of conventional RT-PCR. GII NoV was detected in 88.54% of the samples by real-time NASBA, in 86.46% by TaqMan RT-PCR, in 81.25% by conventional RT-PCR, and in 65.7% by ELISA. The two real-time assays were in agreement for 88.5% of the samples. These results demonstrate that real-time NASBA with a molecular beacon probe is highly sensitive, accurate, and specific for NoV detection in clinical samples. Applying this technique to samples with complex matrix and low viral loads, such as food and environmental samples, could be useful for the detection of NoVs and will improve the prevention of NoV outbreaks.

scholarly journals Development of a recombinase-aided amplification assay for rapid detection of human norovirus GII.4

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Zhiwei Qin ◽  
Liang Xue ◽  
Weicheng Cai ◽  
Junshan Gao ◽  
Yueting Jiang ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Enzyme Reaction ◽  
Detection Sensitivity ◽  
Health Concern ◽  
Clinical Samples ◽  
Nucleic Acid Detection ◽  
Human Norovirus ◽  
Amplification Assay ◽  
Norovirus Gii

Abstract Background Human noroviruses are one of the main causes of foodborne illnesses and represent a serious public health concern. Rapid and sensitive assays for human norovirus detection are undoubtedly necessary for clinical diagnosis, especially in regions without more sophisticated equipment. Method The rapid reverse transcription recombinase-aided amplification (RT-RAA) is a fast, robust and isothermal nucleic acid detection method based on enzyme reaction. This method can complete the sample detection at 39 °C in 30 min. In this study, we successfully established a rapid reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of human norovirus GII.4 and applied this assay to clinical samples, as well as comparison with commercial reverse transcription real-time fluorescence quantitative PCR (RT-qPCR). Results At 95% probability, the detection sensitivity of RT-RAA was 3.425 log10 genomic copies (LGC)/reaction. Moreover, no cross-reaction was observed with other norovirus genogroups and other common foodborne viruses. Stool samples were examined by RT-RAA and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Compared of RT-qPCR, kappa values for human norovirus detection with RT-RAA were 0.894 (p < 0.001), indicating that both assays were in agreement. Conclusion This RT-RAA assay provides a rapid, specific, and sensitive assay for human norovirus detection and is suitable for clinical testing.

scholarly journals SARS-CoV-2 within-host diversity and transmission

Science ◽  
2021 ◽  
Vol 372 (6539) ◽  
pp. eabg0821 ◽  
Author(s):  
Katrina A. Lythgoe ◽  
Matthew Hall ◽  
Luca Ferretti ◽  
Mariateresa de Cesare ◽  
George MacIntyre-Cockett ◽  
...  
Keyword(s):  
United Kingdom ◽  
Severe Acute Respiratory Syndrome ◽  
Phylogenetic Tree ◽  
Immune Escape ◽  
Clinical Samples ◽  
Diversity Patterns ◽  
Host Diversity ◽  
Viral Loads ◽  
The United Kingdom ◽  
Low Levels

Extensive global sampling and sequencing of the pandemic virus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have enabled researchers to monitor its spread and to identify concerning new variants. Two important determinants of variant spread are how frequently they arise within individuals and how likely they are to be transmitted. To characterize within-host diversity and transmission, we deep-sequenced 1313 clinical samples from the United Kingdom. SARS-CoV-2 infections are characterized by low levels of within-host diversity when viral loads are high and by a narrow bottleneck at transmission. Most variants are either lost or occasionally fixed at the point of transmission, with minimal persistence of shared diversity, patterns that are readily observable on the phylogenetic tree. Our results suggest that transmission-enhancing and/or immune-escape SARS-CoV-2 variants are likely to arise infrequently but could spread rapidly if successfully transmitted.

Experimental miniature piglet model for the infection of human norovirus GII

2017 ◽  
Vol 90 (4) ◽  
pp. 655-662 ◽  
Author(s):  
Dong Joo Seo ◽  
Day Jung ◽  
Soontag Jung ◽  
Seung-Kwon Ha ◽  
Sang-Do Ha ◽  
...  
Keyword(s):  
Human Norovirus ◽  
Norovirus Gii

scholarly journals Cattle connection: molecular epidemiology of BVDV outbreaks via rapid nanopore whole-genome sequencing of clinical samples

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Jacqueline King ◽  
Anne Pohlmann ◽  
Kamila Dziadek ◽  
Martin Beer ◽  
Kerstin Wernike
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Economic Losses ◽  
Clinical Samples ◽  
Bovine Viral Diarrhea ◽  
Sequence Information ◽  
Whole Genome ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Loads

Abstract Background As a global ruminant pathogen, bovine viral diarrhea virus (BVDV) is responsible for the disease Bovine Viral Diarrhea with a variety of clinical presentations and severe economic losses worldwide. Classified within the Pestivirus genus, the species Pestivirus A and B (syn. BVDV-1, BVDV-2) are genetically differentiated into 21 BVDV-1 and four BVDV-2 subtypes. Commonly, the 5’ untranslated region and the Npro protein are utilized for subtyping. However, the genetic variability of BVDV leads to limitations in former studies analyzing genome fragments in comparison to a full-genome evaluation. Results To enable rapid and accessible whole-genome sequencing of both BVDV-1 and BVDV-2 strains, nanopore sequencing of twelve representative BVDV samples was performed on amplicons derived through a tiling PCR procedure. Covering a multitude of subtypes (1b, 1d, 1f, 2a, 2c), sample matrices (plasma, EDTA blood and ear notch), viral loads (Cq-values 19–32) and species (cattle and sheep), ten of the twelve samples produced whole genomes, with two low titre samples presenting 96 % genome coverage. Conclusions Further phylogenetic analysis of the novel sequences emphasizes the necessity of whole-genome sequencing to identify novel strains and supplement lacking sequence information in public repositories. The proposed amplicon-based sequencing protocol allows rapid, inexpensive and accessible obtainment of complete BVDV genomes.

scholarly journals Identification of a blockade epitope of human norovirus GII.17

2021 ◽  
pp. 1-17
Author(s):  
Yufang Yi ◽  
Xiaoli Wang ◽  
Shuxia Wang ◽  
Pei Xiong ◽  
Qingwei Liu ◽  
...  
Keyword(s):  
Human Norovirus ◽  
Norovirus Gii

Investigation of Children with Acute Gastroenteritis by Multiplex PCR Method in Central Part of Turkey

2022 ◽  
Author(s):  
Fatih Yılmaz ◽  
Havva Kaya ◽  
Mehmet Özdemir
Keyword(s):  
Multiplex Pcr ◽  
Acute Gastroenteritis ◽  
Hospital Admissions ◽  
Age Groups ◽  
Viral Gastroenteritis ◽  
Viral Agent ◽  
Mortality And Morbidity ◽  
Stool Samples ◽  
Pcr Method ◽  
Norovirus Gii

Abstract Objective Gastroenteritis is a disease that affects all age groups, especially children, and causes high mortality and morbidity in all countries. The most common agents of acute gastroenteritis are viral agents. As a result, millions of diarrhea attacks and hospital admissions occur worldwide every year due to viral gastroenteritis. This study uses the multiplex polymerase chain reaction (PCR) method to investigate the viruses that are the causative agents of viral gastroenteritis in the pediatric patient group in Konya, Turkey. Methods Stool samples of 94 patients aged 0 to 18 years sent from Emergency clinics and Pediatric outpatient clinics, Meram Medical Faculty Hospital Pediatric clinics, Konya Necmettin Erbakan University to Medical Microbiology Laboratory with a diagnosis of gastroenteritis between February and December 2018 were included in the study. Stool samples were stored at –80°C until the time of the analysis. Deoxyribonucleic acid/ribonucleic acid isolation from stool samples was performed with EZ1 Virus Mini Kit v2.0 (Qiagen, Hilden, Germany) using an automatic extraction system (BioRobot EZ1 system, Qiagen). The presence of astrovirus, rotavirus, adenovirus, norovirus (GI, GII), and sapovirus agents was investigated by the multiplex PCR method (Fast Track Diagnostics, Luxembourg) viral gastroenteritis kit. Results Viral gastroenteritis agents were detected in 56.3% of the patients. One viral agent was detected in 47 (50%) of these patients and at least two viral agents in 6 (6.3%) of them. Norovirus GII was detected in 20 (21.2%) of the children included in the study, adenovirus in 13 (13.8%), rotavirus in 11 (12.8%), astrovirus in 11 (11.7%), sapovirus in 4 (4.2%), and norovirus GI in 1 (1.06%). When the distribution of viral agents was examined by months, the most number of agents were observed (21; 35%) in May, followed by April and June (12; 20%). Considering the distribution of the prevalence of the agents by age, it was seen to be mainly between 0 and 12 months (42%). Conclusion Considering that the most common viral agent in our region is norovirus GII, it will be useful to investigate the norovirus that is not routinely examined in children who are admitted to clinics with the complaint of gastroenteritis. It will be appropriate to examine routinely adenovirus, rotavirus, and norovirus in the laboratory, especially in children with diarrhea and vomiting in the winter and spring months.

Transfer of Salmonella and Campylobacter from Stainless Steel to Romaine Lettuce†

2003 ◽  
Vol 66 (12) ◽  
pp. 2231-2236 ◽  
Author(s):  
CHRISTINA M. MOORE ◽  
BRIAN W. SHELDON ◽  
LEE-ANN JAYKUS
Keyword(s):  
Stainless Steel ◽  
Salmonella Typhimurium ◽  
Sampling Period ◽  
Stainless Steel Surface ◽  
Inoculum Level ◽  
Drying Time ◽  
Initial Inoculum ◽  
Significant Difference ◽  
Serovar Typhimurium ◽  
Handling Practices

The degree of transfer of Campylobacter jejuni and Salmonella enterica serovar Typhimurium was evaluated from a stainless steel contact surface to a ready-to-eat food (lettuce). Stainless steel coupons (25 cm2) were inoculated with a 20-μl drop of either C. jejuni or Salmonella Typhimurium to provide an inoculum level of ~106 CFU/28 mm2. Wet and dry lettuce (Lactuca sativa var. longifolia) pieces (9 cm2) were placed onto the inoculated stainless steel surface for 10 s after the designated inoculum drying time (0 to 80 min for C. jejuni; 0 to 120 min for Salmonella Typhimurium), which was followed by the recovery and enumeration of transferred pathogens (lettuce) and residual surface pathogens (stainless steel coupons). For transfers of Salmonella Typhimurium to dry lettuce, there was an increase from 36 to 66% in the percent transfer of the initial inoculum load during the first 60 min of sampling and then a precipitous drop from 66 to 6% in percent transfer. The transfer of Salmonella Typhimurium to wet lettuce ranged from 23 to 31%, with no statistically significant difference between recoveries over the entire 120-min sampling period. For C. jejuni, the mean percent transfer ranged from 16 to 38% for dry lettuce and from 15 to 27% for wet lettuce during the 80-min sampling period. The results of this study indicate that relatively high numbers of bacteria may be transferred to a food even 1 to 2 h after surface contamination. These findings can be used to support future projects aimed at estimating the degree of risk associated with poor handling practices of ready-to-eat foods.

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