Application of Long-range and Binding Reverse Transcription-quantitative PCR to Indicate the Viral Integrities of Noroviruses

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2014 Aug 10

PMID 25107982

Citations 16

Authors

Dan Li

Ann De Keuckelaere

Mieke Uyttendaele

Affiliations

Soon will be listed here.

Abstract

This study intends to establish and apply methods evaluating both viral capsid and genome integrities of human noroviruses (NoVs), which thus far remain nonculturable. Murine norovirus 1 (MNV-1) and human NoV GII.4 in phosphate-buffered saline suspensions were treated with heat, UV light, or ethanol and detected by reverse transcription-quantitative PCR (RT-qPCR), long-range RT-qPCR, binding RT-qPCR, and binding long-range RT-qPCR. For MNV-1 heated at 60°C for 2 and 30 min, limited reductions of genomic copies (<0.3-log) were obtained by RT-qPCR and long-range RT-qPCR, while the cell-binding pretreatments obtained higher reductions (>1.89-log reduction after 60°C for 30 min by binding long-range RT-qPCR). The human NoV GII.4 was found to be more heat resistant than MNV-1. For both MNV-1 and human NoV GII.4 after UV treatments of 20 and 200 mJ/cm(2), no significant difference (P > 0.05) was observed between the dose-dependent reductions obtained by the four detection methodologies. Treatment of 70% ethanol for 1 min was shown to be more effective for inactivation of both MNV-1 and human NoV GII.4 than the heat and UV treatments used in this study. Subsequently, eight raspberry and four shellfish samples previously shown to be naturally contaminated with human NoVs by RT-qPCR (GI and GII; thus, 24 RT-qPCR signals) were subjected to comparison by this method. RT-qPCR, long-range RT-qPCR, binding RT-qPCR, and binding long-range RT-qPCR detected 20/24, 14/24, 24/24, and 23/24 positive signals, respectively, indicating the abundant presence of intact NoV particles.

Citing Articles

Impact of Nanoparticle-Based TiO Surfaces on Norovirus Capsids and Genome Integrity.

Raymond P, St-Germain F, Paul S, Chabot D, Deschenes L Foods. 2024; 13(10).

PMID: 38790828 PMC: 11121413. DOI: 10.3390/foods13101527.

A Multifaceted Approach for Evaluating Hepatitis E Virus Infectivity In Vitro: Cell Culture and Innovative Molecular Methods for Integrity Assessment.

Locus T, Lambrecht E, Lamoral S, Willems S, Van Gucht S, Vanwolleghem T Vet Sci. 2023; 10(12).

PMID: 38133227 PMC: 10748075. DOI: 10.3390/vetsci10120676.

Evaluating Novel Quantification Methods for Infectious Baculoviruses.

Lothert K, Bagrin E, Wolff M Viruses. 2023; 15(4).

PMID: 37112978 PMC: 10141099. DOI: 10.3390/v15040998.

Impact of Capsid and Genomic Integrity Tests on Norovirus Extraction Recovery Rates.

Raymond P, Paul S, Guy R Foods. 2023; 12(4).

PMID: 36832901 PMC: 9957022. DOI: 10.3390/foods12040826.

Investigation on the applicability of a long-range reverse-transcription quantitative polymerase chain reaction assay for the rapid detection of active viruses.

Yasuura M, Nakaya Y, Ashiba H, Fukuda T BMC Microbiol. 2022; 22(1):300.

PMID: 36510144 PMC: 9743722. DOI: 10.1186/s12866-022-02723-7.

References

Li D, Baert L, Xia M, Zhong W, Van Coillie E, Jiang X . Evaluation of methods measuring the capsid integrity and/or functions of noroviruses by heat inactivation. J Virol Methods. 2012; 181(1):1-5. DOI: 10.1016/j.jviromet.2012.01.001. View

Sano D, Pinto R, Omura T, Bosch A . Detection of oxidative damages on viral capsid protein for evaluating structural integrity and infectivity of human norovirus. Environ Sci Technol. 2009; 44(2):808-12. DOI: 10.1021/es9018964. View

Hirneisen K, Black E, Cascarino J, Fino V, Hoover D, Kniel K . Viral Inactivation in Foods: A Review of Traditional and Novel Food-Processing Technologies. Compr Rev Food Sci Food Saf. 2021; 9(1):3-20. DOI: 10.1111/j.1541-4337.2009.00092.x. View

Stals A, Baert L, Botteldoorn N, Werbrouck H, Herman L, Uyttendaele M . Multiplex real-time RT-PCR for simultaneous detection of GI/GII noroviruses and murine norovirus 1. J Virol Methods. 2009; 161(2):247-53. DOI: 10.1016/j.jviromet.2009.06.019. View

Steyer A, Godic Torkar K, Gutierrez-Aguirre I, Poljsak-Prijatelj M . High prevalence of enteric viruses in untreated individual drinking water sources and surface water in Slovenia. Int J Hyg Environ Health. 2011; 214(5):392-8. DOI: 10.1016/j.ijheh.2011.05.006. View

Baert L, Uyttendaele M, Van Coillie E, Debevere J . The reduction of murine norovirus 1, B. fragilis HSP40 infecting phage B40-8 and E. coli after a mild thermal pasteurization process of raspberry puree. Food Microbiol. 2008; 25(7):871-4. DOI: 10.1016/j.fm.2008.06.002. View

Gassilloud B, Duval M, Schwartzbrod L, Gantzer C . Recovery of feline calicivirus infectious particles and genome from water: comparison of two concentration techniques. Water Sci Technol. 2003; 47(3):97-101. View

Rodriguez R, Pepper I, Gerba C . Application of PCR-based methods to assess the infectivity of enteric viruses in environmental samples. Appl Environ Microbiol. 2008; 75(2):297-307. PMC: 2620694. DOI: 10.1128/AEM.01150-08. View

Li D, Baert L, Van Coillie E, Uyttendaele M . Critical studies on binding-based RT-PCR detection of infectious Noroviruses. J Virol Methods. 2011; 177(2):153-9. DOI: 10.1016/j.jviromet.2011.07.013. View

10.

Nishida T, Kimura H, Saitoh M, Shinohara M, Kato M, Fukuda S . Detection, quantitation, and phylogenetic analysis of noroviruses in Japanese oysters. Appl Environ Microbiol. 2003; 69(10):5782-6. PMC: 201174. DOI: 10.1128/AEM.69.10.5782-5786.2003. View

11.

DSouza D, Su X . Efficacy of chemical treatments against murine norovirus, feline calicivirus, and MS2 bacteriophage. Foodborne Pathog Dis. 2009; 7(3):319-26. DOI: 10.1089/fpd.2009.0426. View

12.

Li D, Baert L, De Jonghe M, Van Coillie E, Ryckeboer J, Devlieghere F . Inactivation of murine norovirus 1, coliphage phiX174, and Bacteroides [corrected] fragilis phage B40-8 on surfaces and fresh-cut iceberg lettuce by hydrogen peroxide and UV light. Appl Environ Microbiol. 2010; 77(4):1399-404. PMC: 3067203. DOI: 10.1128/AEM.02131-10. View

13.

Haramoto E, Katayama H, Oguma K, Ohgaki S . Recovery of naked viral genomes in water by virus concentration methods. J Virol Methods. 2007; 142(1-2):169-73. DOI: 10.1016/j.jviromet.2007.01.024. View

14.

Wobus C, Karst S, Thackray L, Chang K, Sosnovtsev S, Belliot G . Replication of Norovirus in cell culture reveals a tropism for dendritic cells and macrophages. PLoS Biol. 2004; 2(12):e432. PMC: 532393. DOI: 10.1371/journal.pbio.0020432. View

15.

Escudero-Abarca B, Rawsthorne H, Goulter R, Suh S, Jaykus L . Molecular methods used to estimate thermal inactivation of a prototype human norovirus: more heat resistant than previously believed?. Food Microbiol. 2014; 41:91-5. DOI: 10.1016/j.fm.2014.01.009. View

16.

Knight A, Li D, Uyttendaele M, Jaykus L . A critical review of methods for detecting human noroviruses and predicting their infectivity. Crit Rev Microbiol. 2012; 39(3):295-309. DOI: 10.3109/1040841X.2012.709820. View

17.

Herbst-Kralovetz M, Radtke A, Lay M, Hjelm B, Bolick A, Sarker S . Lack of norovirus replication and histo-blood group antigen expression in 3-dimensional intestinal epithelial cells. Emerg Infect Dis. 2013; 19(3):431-8. PMC: 3647661. DOI: 10.3201/eid1903.121029. View

18.

Duizer E, Schwab K, Neill F, Atmar R, Koopmans M, Estes M . Laboratory efforts to cultivate noroviruses. J Gen Virol. 2004; 85(Pt 1):79-87. DOI: 10.1099/vir.0.19478-0. View

19.

Baert L, Wobus C, Van Coillie E, Thackray L, Debevere J, Uyttendaele M . Detection of murine norovirus 1 by using plaque assay, transfection assay, and real-time reverse transcription-PCR before and after heat exposure. Appl Environ Microbiol. 2007; 74(2):543-6. PMC: 2223245. DOI: 10.1128/AEM.01039-07. View

20.

Baert L, Mattison K, Loisy-Hamon F, Harlow J, Martyres A, Lebeau B . Review: norovirus prevalence in Belgian, Canadian and French fresh produce: a threat to human health?. Int J Food Microbiol. 2011; 151(3):261-9. DOI: 10.1016/j.ijfoodmicro.2011.09.013. View