MERS-CoV Pseudovirus Neutralization Assay

Pseudoviruses carrying MERS-CoV S protein with or without mutations were produced as described elsewhere [25 (link), 26 (link)]. Briefly, pseudovirus was incubated with a test peptide at graded concentrations at 37°C for 1 hour, followed by addition of the virus/peptide mixture to Huh-7 cells. Cultures were refed with fresh medium 12 hours after infection and incubated for another 48 hours at 37°C, followed by addition of luciferase substrate (Promega). Fluorescence was assessed using a luciferase kit (Promega) and an Ultra 384 luminometer (Tecan).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Channappanavar R., Lu L., Xia S., Du L., Meyerholz D.K., Perlman S, & Jiang S. (2015). Protective Effect of Intranasal Regimens Containing Peptidic Middle East Respiratory Syndrome Coronavirus Fusion Inhibitor Against MERS-CoV Infection. The Journal of Infectious Diseases, 212(12), 1894-1903.

Publication 2015

luciferase substrate Ultra 384 luminometer

Cells Fluorescence Infection Luciferase Mers cov Mutations Peptide Promega S protein Virus

Corresponding Organization : New York Blood Center

Other organizations : University of Iowa

Top 5 similar protocols

Protocol cited in 5 other protocols

Variable analysis

independent variables

Concentration of test peptide

dependent variables

Luciferase activity

control variables

Incubation time (1 hour)
Incubation temperature (37°C)
Virus/peptide mixture addition to Huh-7 cells
Refeeding of culture with fresh medium (12 hours after infection)
Incubation time after refeeding (48 hours at 37°C)
Luciferase substrate addition
Luciferase assay using luminometer

positive control

Not explicitly mentioned

negative control

Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!