Microculture virus titration — a simple colourimetric assay for influenza virus titration
Identifieur interne : 001D11 ( Main/Exploration ); précédent : 001D10; suivant : 001D12Microculture virus titration — a simple colourimetric assay for influenza virus titration
Auteurs : Raphael Levi [Israël] ; Talia Beeor-Tzahar [Israël] ; Ruth Arnon [Israël]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 1995.
English descriptors
- Teeft :
- Allantoic, Allantoic fluid, Allantoic fluids, Assay, Average method, Average value, Beit haemek, Biological industries, Calorimetric assay, Cell growth, Cell viability, Chorioallantoic cavity, Colour intensity, Colourimetric, Colourimetric assay, Control wells, Cutoff value, Different cell cultures, Different days, Embryonated, Embryonated eggs, Full automatization, Good correlation, Graphic presentation, Homogenate, Host system, Humidified incubator, Infection medium, Infectious allantoic fluid, Infectious allantoic fluids, Infectious virus, Infectivity, Influenza, Influenza titration, Influenza viruses, Interferon activity, Large number, Lung homogenate samples, Lung homogenates, Many samples, Mcvt, Mcvt method, Mdck, Mdck cells, Mdck cultures, Mice lung homogenates, Microculture virus titration, Microplate wells, Overlay medium, Plaque, Plaque assay, Plaque formation, Positive control, Positive wells, Proportional infectivity, Respiratory syncytial virus, Samples dilutions, Simple colourimetric assay, Tetrazolium, Tissue cultures, Titration, Titration method, Titration methods, Titre, Viable cells, Viral, Viral concentration, Viral particles, Virological, Virological methods, Virus, Virus dose, Virus sample.
Abstract
Abstract: Influenza antigens can be detected by several well established methods. However, when it is important to determine the titre of infective virions, a bioassay should be employed. The standard and the most widely used tests for influenza infectivity are titration carried out in embryonated hen eggs, or the plaque assay employing tissue culture techniques. A simple colourimetric assay for influenza virus detection and titration is described. Samples of allantoic fluid or mice lung homogenates were used to infect MDCK cultures in microplate wells. After an incubation period, the tetrazolium (MTT) colourimetric assay was used to determine cell viability, and when compared to untreated culture control enabled the detection and titration of several influenza strains. When samples were assayed simultaneously in embryonated eggs and by the MCVT method, good correlation in determined titres was obtained. The availability of an additional method for influenza titration allows more flexibility in the choice of titration method according to the specific needs of the study. Furthermore, this method lends itself to full automatization. Similar procedures should also be applicable to titration of other cytopathic viruses.
Url:
DOI: 10.1016/0166-0934(94)00137-6
Affiliations:
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type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995">1995</date><biblScope unit="volume">52</biblScope><biblScope unit="issue">1–2</biblScope><biblScope unit="page" from="55">55</biblScope><biblScope unit="page" to="64">64</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Allantoic</term><term>Allantoic fluid</term><term>Allantoic fluids</term><term>Assay</term><term>Average method</term><term>Average value</term><term>Beit haemek</term><term>Biological industries</term><term>Calorimetric assay</term><term>Cell growth</term><term>Cell viability</term><term>Chorioallantoic cavity</term><term>Colour intensity</term><term>Colourimetric</term><term>Colourimetric assay</term><term>Control wells</term><term>Cutoff 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formation</term><term>Positive control</term><term>Positive wells</term><term>Proportional infectivity</term><term>Respiratory syncytial virus</term><term>Samples dilutions</term><term>Simple colourimetric assay</term><term>Tetrazolium</term><term>Tissue cultures</term><term>Titration</term><term>Titration method</term><term>Titration methods</term><term>Titre</term><term>Viable cells</term><term>Viral</term><term>Viral concentration</term><term>Viral particles</term><term>Virological</term><term>Virological methods</term><term>Virus</term><term>Virus dose</term><term>Virus sample</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Influenza antigens can be detected by several well established methods. However, when it is important to determine the titre of infective virions, a bioassay should be employed. The standard and the most widely used tests for influenza infectivity are titration carried out in embryonated hen eggs, or the plaque assay employing tissue culture techniques. A simple colourimetric assay for influenza virus detection and titration is described. Samples of allantoic fluid or mice lung homogenates were used to infect MDCK cultures in microplate wells. After an incubation period, the tetrazolium (MTT) colourimetric assay was used to determine cell viability, and when compared to untreated culture control enabled the detection and titration of several influenza strains. When samples were assayed simultaneously in embryonated eggs and by the MCVT method, good correlation in determined titres was obtained. The availability of an additional method for influenza titration allows more flexibility in the choice of titration method according to the specific needs of the study. Furthermore, this method lends itself to full automatization. Similar procedures should also be applicable to titration of other cytopathic viruses.</div></front></TEI><affiliations><list><country><li>Israël</li></country></list><tree><country name="Israël"><noRegion><name sortKey="Levi, Raphael" sort="Levi, Raphael" uniqKey="Levi R" first="Raphael" last="Levi">Raphael Levi</name></noRegion><name sortKey="Arnon, Ruth" sort="Arnon, Ruth" uniqKey="Arnon R" first="Ruth" last="Arnon">Ruth Arnon</name><name sortKey="Beeor Tzahar, Talia" sort="Beeor Tzahar, Talia" uniqKey="Beeor Tzahar T" first="Talia" last="Beeor-Tzahar">Talia Beeor-Tzahar</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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