Journal of Microbiology

Journal Article

Gut Lactobacillus and Probiotics Lactobacillus lactis/rhamnosis Ameliorate Liver Fibrosis in Prevention and Treatment: Sung Min Won , Na Young Lee , Ki&# , Haripriya Gupta , Satya Priya Sharma , Kyung Hwan Kim , Byoung Kook Kim , Hyun Chae Joung , Jin Ju Jeong , Raja Ganesan , Sang Hak Han , Sang Jun Yoon , Dong Joon Kim , Ki Tae Suk; J. Microbiol. 2023;61(2):245-257. Published online February 6, 2023; DOI: https://doi.org/10.1007/s12275-023-00014-y

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The progression and exacerbation of liver fibrosis are closely related to the gut microbiome. It is hypothesized that some probiotics may slow the progression of liver fibrosis. In human stool analysis [healthy group (n = 44) and cirrhosis group (n = 18)], difference in Lactobacillus genus between healthy group and cirrhosis group was observed. Based on human data, preventive and therapeutic effect of probiotics Lactobacillus lactis and L. rhamnosus was evaluated by using four mice fibrosis models. L. lactis and L. rhamnosus were supplied to 3,5-diethoxycarbonyl-1,4-dihydrocollidine or carbon tetrachloride-induced liver fibrosis C57BL/6 mouse model. Serum biochemical measurements, tissue staining, and mRNA expression in the liver were evaluated. The microbiome was analyzed in mouse cecal contents. In the mouse model, the effects of Lactobacillus in preventing and treating liver fibrosis were different for each microbe species. In case of L. lactis, all models showed preventive and therapeutic effects against liver fibrosis. In microbiome analysis in mouse models administered Lactobacillus, migration and changes in the ratio and composition of the gut microbial community were confirmed. L. lactis and L. rhamnosus showed preventive and therapeutic effects on the progression of liver fibrosis, suggesting that Lactobacillus intake may be a useful strategy for prevention and treatment.

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Dingkang Wang, Ruijie Xu, Sha Liu, Xiaomin Sun, Tianxiao Zhang, Lin Shi, Youfa Wang
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Lactobacillus gasseri BNR17 and Limosilactobacillus fermentum ABF21069 Ameliorate High Sucrose-Induced Obesity and Fatty Liver via Exopolysaccharide Production and β-oxidation
Yu Mi Jo, Yoon Ji Son, Seul-Ah Kim, Gyu Min Lee, Chang Won Ahn, Han-Oh Park, Ji-Hyun Yun
Journal of Microbiology.2024; 62(10): 907. CrossRef
Probiotics modulation of the endotoxemic effect on the gut and liver of the lipopolysaccharide challenged mice
Gyan Babu, Banalata Mohanty
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Research reviews and prospects of gut microbiota in liver cirrhosis: a bibliometric analysis (2001–2023)
Xiaofei Zhu, Ziyuan Zhou, Xiaxia Pan
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Xue Li, Xinyi Xu, Sian Tao, Yue Su, Li Wen, Dong Wang, Jibin Liu, Quansheng Feng
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Samantha Roldán-Pérez, Sara Lucía Gómez Rodríguez, José Uriel Sepúlveda-Valencia, Orlando Simón Ruiz Villadiego, María Elena Márquez Fernández, Olga I. Montoya Campuzano, Mónica María Durango-Zuleta
Heliyon.2023; 9(11): e21558. CrossRef
Agrocybe aegerita Polysaccharide Combined with Bifidobacterium lactis Bb-12 Attenuates Aging-Related Oxidative Stress and Restores Gut Microbiota
Xiaoyan Liu, Yanyu Feng, Hongmin Zhen, Lina Zhao, Hongqiang Wu, Bin Liu, Guangsen Fan, Aijun Tong
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Probiotics and liver fibrosis: An evidence-based review of the latest research
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Review

T cell responses to SARS-CoV-2 in humans and animals: Sameer-ul-Salam Mattoo , Jinjong Myoung; J. Microbiol. 2022;60(3):276-289. Published online February 14, 2022; DOI: https://doi.org/10.1007/s12275-022-1624-z

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Abstract

SARS-CoV-2, the causative agent of COVID-19, first emerged in 2019. Antibody responses against SARS-CoV-2 have been given a lot of attention. However, the armamentarium of humoral and T cells may have differing roles in different viral infections. Though the exact role of T cells in COVID-19 remains to be elucidated, prior experience with human coronavirus has revealed an essential role of T cells in the outcomes of viral infections. Moreover, an increasing body of evidence suggests that T cells might be effective against SARS-CoV-2. This review summarizes the role of T cells in mouse CoV, human pathogenic respiratory CoV in general and SARSCoV- 2 in specific.

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Vaccine Strategies Against RNA Viruses: Current Advances and Future Directions
Kuei-Ching Hsiung, Huan-Jung Chiang, Sebastian Reinig, Shin-Ru Shih
Vaccines.2024; 12(12): 1345. CrossRef
Intradermal Fractional ChAdOx1 nCoV-19 Booster Vaccine Induces Memory T Cells: A Follow-Up Study
Ratchanon Sophonmanee, Perawas Preampruchcha, Jomkwan Ongarj, Bunya Seeyankem, Porntip Intapiboon, Smonrapat Surasombatpattana, Supattra Uppanisakorn, Pasuree Sangsupawanich, Sarunyou Chusri, Nawamin Pinpathomrat
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Safety and efficacy of COVID-19 vaccine immunization during pregnancy in 1024 pregnant women infected with the SARS-CoV-2 Omicron virus in Shanghai, China
Hongmei Deng, Yinpeng Jin, Minmin Sheng, Min Liu, Jie Shen, Wei Qian, Gang Zou, Yixin Liao, Tiefu Liu, Yun Ling, Xiaohong Fan
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Differential Impact of Spike Protein Mutations on SARS-CoV-2 Infectivity and Immune Evasion: Insights from Delta and Kappa Variants
Tae-Hun Kim, Sojung Bae, Jinjong Myoung
Journal of Microbiology and Biotechnology.2024; 34(12): 2506. CrossRef
Animal models for COVID-19 and tuberculosis
Björn Corleis, Max Bastian, Donata Hoffmann, Martin Beer, Anca Dorhoi
Frontiers in Immunology.2023;[Epub] CrossRef
A mathematical model and simulation scenarios for T and B cells immune response to severe acute respiratory syndrome-coronavirus-2
L Cuesta-Herrera, F Córdova-Lepe, L Pastenes, A D Arencibia, Y Baldera-Moreno, H A Torres-Mantilla
Journal of Physics: Conference Series.2023; 2516(1): 012007. CrossRef
Distinctive Combinations of RBD Mutations Contribute to Antibody Evasion in the Case of the SARS-CoV-2 Beta Variant
Tae-Hun Kim, Sojung Bae, Sunggeun Goo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2023; 33(12): 1587. CrossRef
Two years of COVID-19 pandemic: where are we now?
Jinjong Myoung
Journal of Microbiology.2022; 60(3): 235. CrossRef
Escape and Over-Activation of Innate Immune Responses by SARS-CoV-2: Two Faces of a Coin
Sameer-ul-Salam Mattoo, Seong-Jun Kim, Dae-Gyun Ahn, Jinjong Myoung
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Development of a T Cell-Based COVID-19 Vaccine Using a Live Attenuated Influenza Vaccine Viral Vector
Irina Isakova-Sivak, Ekaterina Stepanova, Victoria Matyushenko, Sergei Niskanen, Daria Mezhenskaya, Ekaterina Bazhenova, Elena Krutikova, Tatiana Kotomina, Polina Prokopenko, Bogdan Neterebskii, Aleksandr Doronin, Elena Vinogradova, Kirill Yakovlev, Konst
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Journal Articles

Crystal structure of the nuclease and capping domain of SbcD from Staphylococcus aureus: Jinwook Lee , Inseong Jo , Jinsook Ahn , Seokho Hong , Soyeon Jeong , Aeran Kwon , Nam-Chul Ha; J. Microbiol. 2021;59(6):584-589. Published online April 20, 2021; DOI: https://doi.org/10.1007/s12275-021-1012-0

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Abstract

The SbcCD complex is an essential component of the DNA double-strand break (DSB) repair system in bacteria. The bacterial SbcCD complex recognizes and cleaves the DNA ends in DSBs by ATP-dependent endo- and exonuclease activities as an early step of the DNA repair process. SbcD consists of nuclease, capping, and helix-loop-helix domains. Here, we present the crystal structure of a SbcD fragment from Staphylococcus aureus, which contained nuclease and capping domains, at a resolution of 2.9 Å. This structure shows a dimeric assembly similar to that of the corresponding domains of SbcD from Escherichia coli. The S. aureus SbcD fragment exhibited endonuclease activities on supercoiled DNA and exonuclease activity on linear and nicked DNA. This study contributes to the understanding of the molecular basis for how bacteria can resist sterilizing treatment, causing DNA damage.

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Staphylococcus aureus SOS response: Activation, impact, and drug targets
Kaiying Cheng, Yukang Sun, Huan Yu, Yingxuan Hu, Yini He, Yuanyuan Shen
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Prevalence of human Norovirus by genotype in contaminated groundwater in Korea over the last decade (2007–2016): Siwon Lee , Junhyeong Jang , Kyungseon Bae , Wonseok Lee , Hyenmi Chung , Sangjung Park; J. Microbiol. 2018;56(12):926-931. Published online November 27, 2018; DOI: https://doi.org/10.1007/s12275-018-8340-8

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Abstract

This study investigated the occurrence of human Norovirus (HuNoV) by genotype in 1,486 groundwater samples collected from 843 groundwater wells suspected of contamination during 2007–2016, in South Korea. We identified and genotyped 186 HuNoV sequences in 178 HuNoV-positive samples using the RIVM-NoroNet norovirus genotyping tool (NGT) and phylogenetic tree analysis based on RIVM-NoroNet reference sequences. HuNoV GII was more prevalent than GI. The major genotypes detected were HuNoV GII.4 (43.0%), GII.22 (15.6%), GI.5 (10.2%), and GI.1 (8.6%); several genotypes accounted for < 5.0% of all HuNoVs, including GII.17, GI.6, GI.4, GII.6, GI.8, GII.3, GII.13, GI.3, GI.7, GI.2, GI.9, GII.1, GII.8, and GII.10. The prevalence of HuNoVs and number of genotypes detected has drastically decreased over the last decade. HuNoV GII.17, the emerging genotype worldwide including Europe and Asia, appeared in Korean groundwater from 2010, dominated in 2013–2014, and continued to be observed. HuNoV GII.4, the major type occurred last decade from Korean groundwater except 2013–2014, continued to be detected and prevalent similar to HuNoV GII.17 in 2016.

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Development of diagnostic systems for wide range and highly sensitive detection of two waterborne hepatitis viruses from groundwater using the conventional reverse transcription nested PCR assay
Kyung-Seon Bae, Siwon Lee, Jin-Young Lee, Ji-Hye Kim, Youn-Lee Joo, Soo Hyung Lee, Hyen-Mi Chung, Kyung-A You
Journal of Virological Methods.2022; 299: 114344. CrossRef
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Alexey S. Vodop’ianov, Ruslan V. Pisanov, Sergey O. Vodop’ianov, Olga S. Chemisova, Artem A. Gerasimenko, Aleksey K. Noskov, Sergey S. Slis, Svetlana A. Nenadskaya, Anastasia D. Koreneva, Alina V. Kolomoitseva, Evgeny V. Kovalev, Anna R. Litovko, Nina V.
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Prez Verónica Emilse, Victoria Matías, Martínez Laura Cecilia, Giordano Miguel Oscar, Masachessi Gisela, DiCola Guadalupe, Ré Viviana Elizabeth, Paván Jorge Victorio, Colina Rodney, Nates Silvia Viviana, Barril Patricia Angélica
LWT.2021; 144: 111201. CrossRef
Characteristics of Norovirus Food Poisoning Outbreaks in Korea in the 2000s
Jong-Gyu Kim, Joong-Soon Kim, Jeong-Gyoo Kim
Journal of Food Protection.2021; 84(3): 472. CrossRef
Prevalence of emerging torque teno virus (TTV) in drinking water, natural waters and wastewater networks (DWNWWS): A systematic review and meta-analysis of the viral pollution marker of faecal and anthropocentric contaminations
Temitope C. Ekundayo
Science of The Total Environment.2021; 771: 145436. CrossRef
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Antiviral activity of Schizonepeta tenuifolia Briquet against noroviruses via induction of antiviral interferons: Yee Ching Ng , Ye Won Kim , Jeong-Su Lee , Sung Joon Lee , Moon Jung Song; J. Microbiol. 2018;56(9):683-689. Published online August 23, 2018; DOI: https://doi.org/10.1007/s12275-018-8228-7

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Abstract

Human noroviruses are the causative agents of non-bacterial gastroenteritis worldwide. The rapid onset and resolution of disease symptoms suggest that innate immune responses are critical for controlling norovirus infection; however, no effective antivirals are yet available. The present study was conducted to examine the antiviral activities of Schizonepeta tenuifolia Briquet extract (STE) against noroviruses. Treatment of human norovirus replicon-bearing HG23 cells with STE at 5 and 10 mg/ml concentrations resulted in the reduction in the viral RNA levels by 77.2% and 85.9%, respectively. STE had no cytotoxic effects on HG23 cells. Treatment of RAW 264.7 cells infected with murine norovirus 1 (MNV-1), a surrogate virus of human noroviruses, with STE at 10 and 20 μg/ml concentrations resulted in the reduction of viral replication by 58.5% and 84.9%, respectively. STE treatment induced the expression of mRNAs for type I and type II interferons in HG23 cells and upregulated the transcription of interferon-β in infected RAW 264.7 cells via increased phosphorylation of interferon regulatory factor 3, a critical transcription regulator for type I interferon production. These
results
suggest that STE inhibits norovirus replication through the induction of antiviral interferon production during virus replication and may serve as a candidate antiviral substance for treatment against noroviruses.

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Karla D. Passalacqua, Jia Lu, Ian Goodfellow, Abimbola O. Kolawole, Jacob R. Arche, Robert J. Maddox, Kelly E. Carnahan, Mary X. D. O’Riordan, Christiane E. Wobus, Mary K. Estes
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Molecular epidemiology of norovirus in asymptomatic food handlers in Busan, Korea, and emergence of genotype GII.17: Hee Soo Koo , Mi Ok Lee , Pyeong Tae Ku , Su Jeong Hwang , Dong Ju Park , Hyung Suk Baik; J. Microbiol. 2016;54(10):686-694. Published online September 30, 2016; DOI: https://doi.org/10.1007/s12275-016-6312-4

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Abstract

The molecular epidemiology of norovirus infections was studied in food handlers without any symptoms from January to December 2015 in Busan city, Korea. A total of 2,174 fecal specimens from asymptomatic food handlers were analyzed, and 2.3% (49/2,174) were norovirus-positive. Fourteen of 335 samples (4.2%) were positive in January; fifteen of 299 samples (5.0%) in February, and seven of 189 samples (3.7%) in December. However, norovirus was rarely detected in other months. From sequencing analysis, 11 genotypes (five GI and six GII genotypes) were detected. Among the 42 capid gene sequences identified, 14 were from the GI genogroup, while 28 were from the GII genogroup. The most commonly detected genotype was GII.17, comprising 15 (35.7%) of positive samples. From January 2012 to December 2015, 5,138 samples were collected from gastroenteritis patients and outbreaks in Busan. The most detected genotype in 2012, 2013, and 2014 was GII.4 (121, 24, and 12 cases, respectively), but in 2015, GII.17 (25 cases) was the most common. The GII.4 genotype was the major cause of acute gastroenteritis from 2012 to 2014, but the GII.17 genotype became the most prevalent cause in 2015. Continued epidemiological surveillance of GII.17 is needed, together with assessment of the risk of norovirus infection.

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Research Support, Non-U.S. Gov'ts

Function of VP2 Protein in the Stability of the Secondary Structure of Virus-like Particles of Genogroup II Norovirus at Different pH Levels: Function of VP2 Protein in the Stability of NoV VLPs: Yao Lin , Li Fengling , Wang Lianzhu , Zhai Yuxiu , Jiang Yanhua; J. Microbiol. 2014;52(11):970-975. Published online October 3, 2014; DOI: https://doi.org/10.1007/s12275-014-4323-6

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Abstract

VP2 is the minor structural protein of noroviruses (NoV) and may function in NoV particle stability. To determine the function of VP2 in the stability of the NoV particle, we constructed and purified two kinds of virus-like particles (VLPs), namely, VLPs (VP1) and VLPs (VP1+VP2), from Sf9 cells infected with recombinant baculoviruses by using a Bac-to-Bac? baculovirus expression system. The two kinds of VLPs were treated with different phosphate buffers (pH 2 to pH 8); the secondary structure was then analyzed by far UV circular dichroism (CD) spectroscopy. Results showed that significant disruptions of the secondary structure of proteins were not observed at pH 2 to pH 7. At pH 8, the percentages of α-helix, β-sheet, and β-turn in VLPs (VP1) were decreased from 11% to 8%, from 37% to 32%, and from 20% to 16%, respectively. The percentage of coil was increased from 32% to 44%. By contrast, the percentages of α-helix, β-sheet, and β-turn in VLPs (VP1+VP2) were decreased from 11% to 10%, from 37% to 35%, and from 20% to 19%, respectively. The percentage of coil was increased from 32% to 36%. VLPs (VP1+VP2) was likely more stable than VLPs (VP1), as indicated by the percentage of the secondary structures analyzed by CD. These results suggested that VP2 could stabilize the secondary structure of VLPs under alkaline pH conditions. This study provided novel insights into the molecular mechanism of the function of VP2 in the stability of NoV particles.

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Genetic Analysis of the Capsid Region of Norovirus GII.4 Variants Isolated in South Korea: Ju-Eun Kim , Sung-Geun Lee , Han-Gil Cho , Sang-Ha Han , Lae-Hyung Kang , Youn-Mi Lee , Chul-Jong Park , Soon-Young Paik; J. Microbiol. 2014;52(5):427-434. Published online April 11, 2014; DOI: https://doi.org/10.1007/s12275-014-3538-x

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Abstract

Norovirus is one of the major causes of non-bacterial gas-troenteritis in humans. The aim of this study was to analyze the amino acid variation of open reading frame 2 of GII.4 variants in South Korea during the period from November 2006 to December 2012. Sixty-nine complete nucleotide se-quences of open reading frame 2 were obtained from 113 GII.4 strains. The GII.4 2006b variants were detected pre-dominantly between 2006 and 2009; however, new GII.4 variants, which were termed the 2010 variant and the 2012 variant, emerged in 2010 and 2012, respectively. The num-ber of GII.4 2006b variants steadily decreased until 2012, whereas the number of gastroenteritis cases caused by the new variants increased between 2010 and 2012. The amino acid sequence in the ORF2 region obtained in this study was compared with other GII.4 variants isolated in various countries. Amino acid variations were observed primarily at epitope sites and the surrounding regions. Amino acids 294, 359, 393, and 413 of the P2 subdomain were the most variable sites among the GII.4 variants. The information in this study can be useful in basic research to predict the emergence and determine the genetic functions of new GII.4 variants.

Citations

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Determining the efficacy of 27 commercially available disinfectants against human noroviruses
Jae-Woong Lee, Lae-Hyung Kang, Min-Kyeong Kim, Jeong-Soon Kim, Myung L. Kim, Sung-Geun Lee, In-Hye Choi, Chul-Jong Park, Soon-Young Paik
Journal of Infection and Public Health.2021; 14(2): 244. CrossRef
Evolutionary changes in the capsid P2 region of Australian strains of the norovirus GII.Pe_GII.4
Leesa D. Bruggink, Jean M. Moselen, Jason A. Roberts, John A. Marshall
Journal of Medical Microbiology.2017; 66(7): 1014. CrossRef
A norovirus intervariant GII.4 recombinant in Victoria, Australia, June 2016: the next epidemic variant?
Leesa Bruggink, Michael Catton, John Marshall
Eurosurveillance.2016;[Epub] CrossRef
Complete Nucleotide Sequence Analysis of the Norovirus GII.4 Sydney Variant in South Korea
Ji-Sun Park, Sung-Geun Lee, Ji-Young Jin, Han-Gil Cho, Weon-Hwa Jheong, Soon-Young Paik
BioMed Research International.2015; 2015: 1. CrossRef
Molecular epidemiology of norovirus GII.4 variants in children under 5 years with sporadic acute gastroenteritis in South Korea during 2006–2013
Han-Gil Cho, Sung-Geun Lee, Ju-Eun Kim, Kyeong-Sin Yu, Deog-Yong Lee, Po-Hyun Park, Mi-hye Yoon, Eek-Hoon Jho, Jaehong Kim, Soon-Young Paik
Journal of Clinical Virology.2014; 61(3): 340. CrossRef

Reliability of Non-Culturable Virus Monitoring by PCR-Based Detection Methods in Environmental Waters Containing Various Concentrations of Target RNA: Eung Seo Koo , Chang-Hoon Yoo , Youjin Na , Soo Young Park , Hey Rhyoung Lyoo , Yong Seok Jeong; J. Microbiol. 2012;50(5):726-734. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2279-y

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9 Scopus

Abstract: Owing to the lack of practical cell culture system for human noroviruses (HuNoV), various detection methods based on conventional reverse transcription-PCR (RT-PCR) and the quantitative real-time PCR have been major tools for monitoring environmental water safety. In this study, we showed that the proportion of water sample concentrates used for one-step RT-PCR significantly influences false-negative findings of the non-culturable viruses. In total, 59 archived samples of previously analyzed water concentrates were reexamined for HuNoV RNA by the one-step RT-PCR and semi-nested PCR. Using new aliquots for RNA extraction for every trial, up to 20 PCR trials were performed for each archive to determine whether the crosscheck results supported the previous determinations. We reconfirmed that 27.6% (8/29) of the samples were HuNoV-positive samples: 6.7% (1/15) from groundwater, 33.3% (3/9) from river water, and 80% (4/5) from treated sewage effluent (TSE). These results corresponded to the ratio of previously negative HuNoV samples now identified as positive (8/30): 6.7% (1/15) from groundwater, 20% (1/5) from river water, and 60% (6/10) from TSE. To elucidate the cause of these results, 16 different concentrations of murine norovirus (MNV) RNA (from 2×102 to 8×103 copies, divided into 10 tubes for each concentration) were subjected to one-step RT-PCR. The detection frequency and reproducibility decreased sharply when the number of MNV RNA copies fell below threshold levels. These observations suggest that the proportion of water concentrate used for PCR-based detection should be considered carefully when deciding viral presence in certain types of environmental water, particularly in regard with legal controls.

cDNA Cloning of Korean Human Norovirus and Nucleotidylylation of VPg by Norovirus RNA-Dependent RNA Polymerase: Byung Sup Min , Kang Rok Han , Jung Ihn Lee , Jai Myung Yang; J. Microbiol. 2012;50(4):625-630. Published online August 25, 2012; DOI: https://doi.org/10.1007/s12275-012-2087-4

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4 Scopus

Abstract: Norovirus, a member of the Caliciviridae family, is a major causative agent of gastroenteritis worldwide. The cDNA of the entire genome of human norovirus (HuNV) was cloned using the RNA extracted from the stool sample of a Korean patient. The RNA genome consists of 7,559 nucleotides, carries 3 open reading frames (ORFs), 5' and 3' noncoding regions, and a poly(A) tail at the 3' end. Phylogenic analysis of the nucleotide sequence indicated that it belongs to GII.4, the most dominant genogroup. To analyze RNA synthesis and nucleotidylylation of VPg by RNA-dependent RNA polymerase (RdRp), recombinant RdRp and VPg were expressed in Escherichia coli as His-tagged forms. The HuNV RdRp exhibited template and divalent cation-dependent RNA synthesis in vitro. The HuNV RdRp nucleotidylylated HuNV VPg but not murine norovirus (MNV) VPg, whereas MNV RdRp nucleotidylylated both MNV and HuNV VPg more efficiently than HuNV RdRp.

Evaluation of the Sensitivity and Specificity of Primer Pairs and the Efficiency of RNA Extraction Procedures to Improve Noroviral Detection from Oysters by Nested Reverse Transcription-Polymerase Chain Reaction: Cheonghoon Lee , Sooryun Cheong , Hee-Jung Lee , Miye Kwon , Ilnam Kang , Eun-Gyoung Oh , Hong-Sik Yu , Soon-Bum Shin , Sang-Jong Kim; J. Microbiol. 2010;48(5):586-593. Published online November 3, 2010; DOI: https://doi.org/10.1007/s12275-010-0047-4

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6 Scopus

Abstract: Noroviruses (NoV) are the key cause of acute epidemic gastroenteritis, and oysters harvested from NoVpolluted sea areas are considered as the significant vectors of viral transmission. To improve NoV detection from oyster using nested reverse transcription-polymerase chain reaction (RT-PCR), we evaluated the sensitivity and specificity of previously published primer pairs and the efficiency of different RNA extraction procedures. Among the primer pairs used for RT-PCR, the sensitivity of GIF1/GIR1-GIF2/GIR1 and GIIF1/GIIR1-GIIF2/GIIR1 was higher than that of other primer pairs used in nested RT-PCR for the detection of NoV genogroup I (NoV GI) and NoV GII from both NoV-positive stool suspension and NoVseeded oyster concentrates, respectively; the resulting products showed neither unspecific bands in the positive samples nor false-positive bands in the negative controls. The extraction of NoV RNA from oyster samples using a QIAamp? Viral RNA Mini kit with a QIAshredderTM Homogenizer pretreatment afforded more efficient recovery (mean recovery for NoV GI and GII, 6.4%) and the procedure was less time consuming (<30 min) than most other RNA extraction procedures. The results of RNA extraction procedure and primer pairs evaluated by nested RT-PCR assay in this study can be useful for monitoring NoV contamination in oysters, which is an indicator of possible public health risks.

Development of a Latex Agglutination Test for Norovirus Detection: Heetae Lee , YoungBin Park , Misoon Kim , Youngmee Jee , Doo-sung Cheon , Hae Sook Jeong , GwangPyo Ko; J. Microbiol. 2010;48(4):419-425. Published online August 20, 2010; DOI: https://doi.org/10.1007/s12275-010-0071-4

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9 Scopus

Abstract: Norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Currently, reverse transcription polymerase chain reaction (RT-PCR) is used commonly to detect NoVs in both clinical and environmental samples. However, RT-PCR requires expensive equipment and cannot be performed on site. In this study, a latex agglutination test (LAT) using antibody-labeled latex beads for detecting NoVs was developed. Two kinds of polyclonal antibodies, one generated from synthetic peptides and the other from E. coli-expressed NoV capsid proteins, were used to develop the LAT. Each of these polyclonal antibodies was immobilized on the surface of latex beads and tested for the ability to detect NoVs. Under optimized conditions, our LAT detected GII.4 NoV at concentrations as low as 3.3×105 RT-PCR units/ml in stool samples. The detection limit for the LAT was approximately 1.7×103 RT-PCR units. Forty-eight stool samples were tested for NoVs using this LAT. In comparison with an RT-PCR assay, the sensitivity and specificity of the LAT were 35% and 100%, respectively. With further optimization, this LAT used with appropriate antibodies could be applied for convenient detection of NoVs in clinical diagnosis and food monitoring.

Development of a Virus Concentration Method and its Application for the Detection of Noroviruses in Drinking Water in China: Junyi Liu , Qingping Wu , Xiaoxia Kou; J. Microbiol. 2007;45(1):48-52.; DOI: https://doi.org/2492 [pii]

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Abstract: A new procedure for the concentration of nonoviruses from water samples has been developed. This procedure (calcium flocculation-citrate dissolution method) uses the following steps: virus flocculation formed by treatment with 1 M CaCl2 and 1 M Na2HPO4, virus release by sodium citrate dissolution (0.3 M Na citrate, pH 3.5), and virus re-concentration by ultrafiltration. When reverse transcription (RT)-PCR was performed after the procedure, the overall detection sensitivity for seeded noroviruses in a one liter drinking water sample was as low as 1 RT-PCR unit, which is equal to a 10-6 dilution of the virus sample. This approach showed at least a 5-fold-higher sensitivity than the current method with its three steps of adsorption-elution-concentration. The newly developed procedure was used to test different brands of bottled drinking water from China for putative contamination with noroviruses. A total of 144 samples were analyzed; all of the samples were negative for norovirus specific nucleic acids.

Validation Study

Rapid Detection of Noroviruses in Fecal Samples and Shellfish by Nucleic Acid Sequence-based Amplification: Xiaoxia Kou , Qingping Wu , Jumei Zhang , Hongying Fan; J. Microbiol. 2006;44(4):403-408.; DOI: https://doi.org/2413 [pii]

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Abstract: The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (NASBA) method of detecting noroviruses in artificially and naturally contaminated shellfish. We used 58 fecal samples that tested positive for noroviruses with electron microscopy (EM) to develop an NASBA assay for these viruses. Oligonucleotide primers targeting the polymerase coding region were used to amplify the viral RNA in an isothermal process that resulted in the accumulation of RNA amplicons. These amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup I (GI) and genogroup II (GII) of noroviruses. The expected band of 327 bp appeared in denaturing agarose gel without any nonspecific band. The specific signal for each amplicon was obtained through Northern blotting in many repeats. All fecal samples of which 46 (79.3%) belonged to GII and 12 (20.6%) belonged to GI were positive for noroviruses by EM and by NASBA. Target RNA concentrations as low as 5 pg/ml were detected in fecal specimens using NASBA. When the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100 pg/1.5 g shellfish tissue. The potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in Guangzhou city of China, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6%) belonged to GII and 5 (20.4%) belonged to GI. The NASBA assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity.

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