Plaque assay method for arboviruses in mosquito cells

1979 ◽  
Vol 5 (2) ◽  
pp. 1077-1080 ◽  
Author(s):  
Conrad E. Yunker ◽  
Jack Cory
Keyword(s):  
Plaque Assay ◽  
Assay Method ◽  
Mosquito Cells

A simple plaque assay method for antibody-dependent cell-mediated cytotoxicity using Cunningham's chamber

1981 ◽  
Vol 42 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Jo Satoh ◽  
Noboru Suzuki ◽  
Yoshio Gotoh ◽  
Katsuo Kumagai
Keyword(s):  
Plaque Assay ◽  
Assay Method ◽  
Dependent Cell

Mechanism of Action of the Anti-Influenza Virus Active Kampo (Traditional Japanese Herbal) Medicine, Hochuekkito

Pharmacology ◽  
2017 ◽  
Vol 101 (3-4) ◽  
pp. 148-155 ◽  
Author(s):  
Katsuaki Dan ◽  
Keita Takanashi ◽  
Hiroko Akiyoshi ◽  
Kaori Munakata ◽  
Hideki Hasegawa ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Mechanism Of Action ◽  
Plaque Assay ◽  
Virus Titer ◽  
Acid Synthesis ◽  
Infection Process ◽  
Assay Method ◽  
Virus Adsorption ◽  
Virus Rna ◽  
Viral Particles

When the Kampo medicine, Hochuekkito (Hochu), was administered to normal mice for 2 weeks, influenza virus titer was reduced. The mechanism of action of Hochu was examined using the plaque assay method. It was suggested that Hochu may either obstruct the first stage of the infection process (adsorption and entry) or may directly target viral particles. Using the plaque assay method, these 2 modes of action could not be differentiated. Virus RNA in the infected cell was verified by quantitative real-time polymerase chain reaction. An equal inhibition effect was obtained when Hochu was preprocessed for normal cells and when they were made to act simultaneously with virus adsorption. The viral load at the cell surface following UV irradiation was higher in the Hochu-administered group as compared with that of the control. Moreover, the affinity of Hochu for the influenza virus was hundred times higher than its affinity for the host cell. The effect of entry obstruction by Hochu was observed via image analysis, where the amount of virus nucleocapsid protein (NP) invading the cell was visualized with FITC-labeled NP antibody. Hochu does not seem to have an effect on nucleic acid synthesis, viral release from infected cells, and on the subsequent second round of infection. In conclusion, Hochu binds to viral particles and forms complexes that can obstruct the entry of influenza virus into cells.

scholarly journals Plaque assay of neonatal calf diarrhea virus and the neutralizing antibody in human sera

1977 ◽  
Vol 5 (1) ◽  
pp. 1-4
Author(s):  
S Matsuno ◽  
S Inouye ◽  
R Kono
Keyword(s):  
Plaque Assay ◽  
Neutralizing Antibody ◽  
Assay Method ◽  
Kidney Cell Line ◽  
Diarrhea Virus ◽  
Bovine Rotavirus ◽  
Neonatal Calf ◽  
Calf Diarrhea ◽  
Human Sera ◽  
Neonatal Calf Diarrhea

Neonatal calf diarrhea virus (a bovine rotavirus) formed distinct plaques in monolayers of MA-104 cells, an established macacus rhesus monkey kidney cell line, when diethylaminoethyl dextran and trypsin were included in the overlay medium. By using this plaque assay method, titration of neutralizing antibody to neonatal calf diarrhea virus was made feasible. It was demonstrated that some human sera contained neutralizing antibody to this agent.

scholarly journals Decanoyl-Arg-Val-Lys-Arg-Chloromethylketone: An Antiviral Compound That Acts against Flaviviruses through the Inhibition of Furin-Mediated prM Cleavage

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1011 ◽  
Author(s):  
Imran ◽  
Saleemi ◽  
Chen ◽  
Wang ◽  
Zhou ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Mammalian Cells ◽  
Plaque Assay ◽  
Virus Titer ◽  
Vero Cells ◽  
Immunofluorescence Assay ◽  
Viral Rna ◽  
Antiviral Compound ◽  
Mosquito Cells ◽  
Infection Treatment

Flaviviruses, such as Zika virus (ZIKV), Japanese encephalitis virus (JEV), Dengue virus (DENV), and West Nile virus (WNV), are important arthropod-borne pathogens that present an immense global health problem. Their unpredictable disease severity, unusual clinical features, and severe neurological manifestations underscore an urgent need for antiviral interventions. Furin, a host proprotein convertase, is a key contender in processing flavivirus prM protein to M protein, turning the inert virus to an infectious particle. For this reason, the current study was planned to evaluate the antiviral activity of decanoyl-Arg-Val-Lys-Arg-chloromethylketone, a specific furin inhibitor, against flaviviruses, including ZIKV and JEV. Analysis of viral proteins revealed a significant increase in the prM/E index of ZIKV or JEV in dec-RVKR-cmk-treated Vero cells compared to DMSO-treated control cells, indicating dec-RVKR-cmk inhibits prM cleavage. Plaque assay, qRT-PCR, and immunofluorescence assay revealed a strong antiviral activity of dec-RVKR-cmk against ZIKV and JEV in terms of the reduction in virus progeny titer and in viral RNA and protein production in both mammalian cells and mosquito cells. Time-of-drug addition assay revealed that the maximum reduction of virus titer was observed in post-infection treatment. Furthermore, our results showed that dec-RVKR-cmk exerts its inhibitory action on the virus release and next round infectivity but not on viral RNA replication. Taken together, our study highlights an interesting antiviral activity of dec-RVKR-cmk against flaviviruses.

Optimal conditions for titration of SV40 by the plaque assay method

1983 ◽  
Vol 7 (2) ◽  
pp. 93-102 ◽  
Author(s):  
James L. Fendrick ◽  
Lesley M. Hallick
Keyword(s):  
Plaque Assay ◽  
Assay Method ◽  
Optimal Conditions

scholarly journals A Simple Methodological Approach for Counting and Identifying Culturable Viruses Adsorbed to Cellulose Nitrate Membrane Filters

2000 ◽  
Vol 66 (1) ◽  
pp. 194-198 ◽  
Author(s):  
Georgios T. Papageorgiou ◽  
Laura Mocé-Llivina ◽  
Christina G. Christodoulou ◽  
Francisco Lucena ◽  
Dina Akkelidou ◽  
...  
Keyword(s):  
Tap Water ◽  
Membrane Filter ◽  
Methodological Approach ◽  
Plaque Assay ◽  
Cellulose Nitrate ◽  
Assay Method ◽  
Most Probable Number ◽  
Cellulose Membrane ◽  
Probable Number ◽  
Membrane Filters

ABSTRACT We identified conditions under which Buffalo green monkey cells grew on the surfaces of cellulose nitrate membrane filters in such a way that they covered the entire surface of each filter and penetrated through the pores. When such conditions were used, poliovirus that had previously been adsorbed on the membranes infected the cells and replicated. A plaque assay method and a quantal method (most probable number of cytopathic units) were used to detect and count the viruses adsorbed on the membrane filters. Polioviruses in aqueous suspensions were then concentrated by adsorption to cellulose membrane filters and were subsequently counted without elution, a step which is necessary when the commonly used methods are employed. The pore size of the membrane filter, the sample contents, and the sample volume were optimized for tap water, seawater, and a 0.25 M glycine buffer solution. The numbers of viruses recovered under the optimized conditions were more than 50% greater than the numbers counted by the standard plaque assay. When ceftazidime was added to the assay medium in addition to the antibiotics which are typically used, the method could be used to study natural samples with low and intermediate levels of microbial pollution without decontamination of the samples. This methodological approach also allowed plaque hybridization either directly on cellulose nitrate membranes or on Hybond N+ membranes after the preparations were transferred.

scholarly journals STUDIES OF THE FATE OF TYPE 1 POLIOVIRUSES IN FLIES

1961 ◽  
Vol 113 (1) ◽  
pp. 159-176 ◽  
Author(s):  
Margrét G. Gudnadóttir
Keyword(s):  
Room Temperature ◽  
Plaque Assay ◽  
Common Species ◽  
Relative Increase ◽  
Assay Method ◽  
Twofold Increase

Studies on the fate of type 1 polioviruses in two common species of flies were carried out. The amount of virus in carcasses and excreta at different times was determined by the plaque assay method. Flies and their excreta remained infective for 11 days when kept at room temperature or when incubated at 36°C. for 2 hours a day. Flies remained infective for 3 months when kept in hibernation. A relative increase in titer was found to occur between 9 and 18 hours after feeding if the flies were incubated at 36°C. for 5 to 15 hours a day. The peak occurred later, at 40 to 52 hours after feeding if less incubation was used. Titers in excreta were parallel to titers in carcasses. A twofold increase in titer over the initial feeding was observed on 3 occasions with type 1 Mahoney but not with the LSc strain of virus.

IMMUNOFLUORESCENCE IN THE STUDY OF VIRUSES IN TISSUE CULTURE: I. DEVELOPMENT OF AN IMMUNOFLUORESCENT FOCUS ASSAY FOR HERPES SIMPLEX VIRUS

1967 ◽  
Vol 13 (11) ◽  
pp. 1489-1497 ◽  
Author(s):  
S. A. Sattar ◽  
J. C. N. Westwood
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Direct Relationship ◽  
Plaque Assay ◽  
Assay Method ◽  
Virus Concentration ◽  
Kidney Cells ◽  
Sensitive Assay ◽  
Host System ◽  
Simplex Virus

A rapid and sensitive assay method has been developed for herpes simplex virus in kidney cells of Cercopithecus aethiops. Using immunofluorescence, foci of infection produced by this virus can be scored within 20–22 hours after inoculation. It has been demonstrated that this procedure gives reproducible results and exhibits a direct relationship between virus concentration and the number of fluorescent foci. The technique is slightly more sensitive than the conventional plaque assay method using the same host system.

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