Development of a Fluorescent in Situ Method for Visualization of Enteric Viruses

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2009 Oct 27

PMID 19854924

Citations 10

Authors

Helen Rawsthorne

Trevor G Phister

Lee-Ann Jaykus

Affiliations

Soon will be listed here.

Abstract

Studying the interactions between enteric pathogens and their environment is important to improving our understanding of their persistence and transmission. However, this remains challenging in large part because of difficulties associated with tracking pathogens in their natural environment(s). In this study, we report a fluorescent labeling strategy which was applied to murine norovirus (MNV-1), a human norovirus surrogate, and hepatitis A virus (HAV). Specifically, streptavidin-labeled Quantum dots (Q-Dots) were bound to biotinylated capsids of MNV-1 and HAV (bio-MNV-1 and bio-HAV); the process was confirmed by using a sandwich-type approach in which streptavidin-bound plates were reacted with biotinylated virus followed by a secondary binding to Q-Dots with an emission range of 635 to 675 nm (Q-Dots 655). The assay demonstrated a relative fluorescence of 528 +/- 48.1 and 112 +/- 8.6 for bio-MNV-1 and control MNV-1, respectively. The biotinylation process did not impact virus infectivity, nor did it interfere with the interactions between the virus and host cells or model produce items. Using fluorescent microscopy, it was possible to visualize both bio-HAV and bio-MNV-1 attached to the surfaces of permissive mammalian cells and green onion tissue. The method provides a powerful tool for the labeling and detection of enteric viruses (and their surrogates) which can be used to track virus behavior in situ.

Citing Articles

Application of an Optimized Direct Lysis Method for Viral RNA Extraction Linking Contaminated Dates to Infection With Hepatitis A Virus.

Rajiuddin S, Midgley S, Jensen T, Muller L, Schultz A Front Microbiol. 2020; 11:516445.

PMID: 33042044 PMC: 7522280. DOI: 10.3389/fmicb.2020.516445.

An Optimised Direct Lysis Method for Viral RNA Extraction and Detection of Foodborne Viruses on Fruits and Vegetables.

Rajiuddin S, Jensen T, Hansen T, Schultz A Food Environ Virol. 2020; 12(3):226-239.

PMID: 32651775 DOI: 10.1007/s12560-020-09437-x.

Evaluation of Methods for the Concentration and Extraction of Viruses from Sewage in the Context of Metagenomic Sequencing.

Hjelmso M, Hellmer M, Fernandez-Cassi X, Timoneda N, Lukjancenko O, Seidel M PLoS One. 2017; 12(1):e0170199.

PMID: 28099518 PMC: 5242460. DOI: 10.1371/journal.pone.0170199.

Development of a nucleic Acid extraction procedure for simultaneous recovery of DNA and RNA from diverse microbes in water.

Hill V, Narayanan J, Gallen R, Ferdinand K, Cromeans T, Vinje J Pathogens. 2015; 4(2):335-54.

PMID: 26016775 PMC: 4493477. DOI: 10.3390/pathogens4020335.

Fluorosomes: fluorescent virus-like nanoparticles that represent a convenient tool to visualize receptor-ligand interactions.

Wojta-Stremayr D, Pickl W Sensors (Basel). 2013; 13(7):8722-49.

PMID: 23881135 PMC: 3758619. DOI: 10.3390/s130708722.

References

Le Guyader F, Mittelholzer C, Haugarreau L, Hedlund K, Alsterlund R, Pommepuy M . Detection of noroviruses in raspberries associated with a gastroenteritis outbreak. Int J Food Microbiol. 2004; 97(2):179-86. DOI: 10.1016/j.ijfoodmicro.2004.04.018. View

Ootsubo M, Shimizu T, Tanaka R, Sawabe T, Tajima K, Yoshimizu M . Oligonucleotide probe for detecting Enterobacteriaceae by in situ hybridization. J Appl Microbiol. 2002; 93(1):60-8. DOI: 10.1046/j.1365-2672.2002.01668.x. View

Parak W, Pellegrino T, Plank C . Labelling of cells with quantum dots. Nanotechnology. 2011; 16(2):R9-R25. DOI: 10.1088/0957-4484/16/2/R01. View

Chan W, Nie S . Quantum dot bioconjugates for ultrasensitive nonisotopic detection. Science. 1998; 281(5385):2016-8. DOI: 10.1126/science.281.5385.2016. View

Arya H, Kaul Z, Wadhwa R, Taira K, Hirano T, Kaul S . Quantum dots in bio-imaging: Revolution by the small. Biochem Biophys Res Commun. 2005; 329(4):1173-7. DOI: 10.1016/j.bbrc.2005.02.043. View

Wheeler C, Vogt T, Armstrong G, Vaughan G, Weltman A, Nainan O . An outbreak of hepatitis A associated with green onions. N Engl J Med. 2005; 353(9):890-7. DOI: 10.1056/NEJMoa050855. View

Volkin D, Burke C, Marfia K, Oswald C, Wolanski B, Middaugh C . Size and conformational stability of the hepatitis A virus used to prepare VAQTA, a highly purified inactivated vaccine. J Pharm Sci. 1997; 86(6):666-73. DOI: 10.1021/js960475h. View

Kampani K, Quann K, Ahuja J, Wigdahl B, Khan Z, Jain P . A novel high throughput quantum dot-based fluorescence assay for quantitation of virus binding and attachment. J Virol Methods. 2007; 141(2):125-32. PMC: 1975807. DOI: 10.1016/j.jviromet.2006.11.043. View

Showell D, Sundkvist T, Reacher M, Gray J . Norovirus outbreak associated with canteen salad in Suffolk, United Kingdom. Euro Surveill. 2007; 12(11):E071129.6. DOI: 10.2807/esw.12.48.03323-en. View

10.

Fang Q, Brockmann S, Botzenhart K, Wiedenmann A . Improved detection of Salmonella spp. in foods by fluorescent in situ hybridization with 23S rRNA probes: a comparison with conventional culture methods. J Food Prot. 2003; 66(5):723-31. DOI: 10.4315/0362-028x-66.5.723. View

11.

Skulstad S, Rodahl E, Jakobsen K, Langeland N, Haarr L . Labeling of surface proteins of herpes simplex virus type 1 using a modified biotin-streptavidin system. Virus Res. 1995; 37(3):253-70. DOI: 10.1016/0168-1702(95)00036-p. View

12.

Bidawid S, Farber J, Sattar S, Hayward S . Heat inactivation of hepatitis A virus in dairy foods. J Food Prot. 2000; 63(4):522-8. DOI: 10.4315/0362-028x-63.4.522. View

13.

Hutin Y, Pool V, CRAMER E, Nainan O, Weth J, Williams I . A multistate, foodborne outbreak of hepatitis A. National Hepatitis A Investigation Team. N Engl J Med. 1999; 340(8):595-602. DOI: 10.1056/NEJM199902253400802. View

14.

Papafragkou E, Plante M, Mattison K, Bidawid S, Karthikeyan K, Farber J . Rapid and sensitive detection of hepatitis A virus in representative food matrices. J Virol Methods. 2007; 147(1):177-87. DOI: 10.1016/j.jviromet.2007.08.024. View

15.

Schmid D, Stuger H, Lederer I, Pichler A, Kainz-Arnfelser G, Schreier E . A foodborne norovirus outbreak due to manually prepared salad, Austria 2006. Infection. 2007; 35(4):232-9. DOI: 10.1007/s15010-007-6327-1. View

16.

Chancellor D, Tyagi S, Bazaco M, Bacvinskas S, Chancellor M, Dato V . Green onions: potential mechanism for hepatitis A contamination. J Food Prot. 2006; 69(6):1468-72. DOI: 10.4315/0362-028x-69.6.1468. View

17.

Harders J, Lukacs N, Jovin T, Riesner D . Imaging of viroids in nuclei from tomato leaf tissue by in situ hybridization and confocal laser scanning microscopy. EMBO J. 1989; 8(13):3941-9. PMC: 401569. DOI: 10.1002/j.1460-2075.1989.tb08577.x. View

18.

Hjertqvist M, Johansson A, Svensson N, Abom P, Magnusson C, Olsson M . Four outbreaks of norovirus gastroenteritis after consuming raspberries, Sweden, June-August 2006. Euro Surveill. 2006; 11(9):E060907.1. DOI: 10.2807/esw.11.36.03038-en. View

19.

Cannon J, Papafragkou E, Park G, Osborne J, Jaykus L, Vinje J . Surrogates for the study of norovirus stability and inactivation in the environment: aA comparison of murine norovirus and feline calicivirus. J Food Prot. 2006; 69(11):2761-5. DOI: 10.4315/0362-028x-69.11.2761. View