Comparison of VSV Pseudovirus and Focus Reduction Neutralization Assays for Measurement of Anti-Andes orthohantavirus Neutralizing Antibodies in Patient Samples

Vial, Cecilia; Whitaker, Annalis; Wilhelm, Jan; Ovalle, Jimena; Perez, Ruth M; Valdivieso, Francisca; Ferrés, Marcela; Martínez-Valdebenito, Constanza; Eisenhauer, Philip; Mertz, Grégory; Hooper, Jay W.; Botten, Jason; Víal, Pablo A.

doi:10.3389/fcimb.2020.00444

Cited by 5 publications

(3 citation statements)

References 17 publications

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“…The neutralizing antibody detection method based on pseudovirus primarily uses HIV, VSV, and other vectors to package viral. From the perspective of biosafety, most of the packaging vectors used presently produce replication-deficient recombinant virus; that is, the virus particles obtained after packaging cannot be amplified in vitro and need to be “real-time rescue “brings experimental and economic pressure to relevant research, ( Whitt, 2010 ; Vial et al, 2020 ; Salazar-Garcia et al, 2021 ). At the same time, a luciferase reporter gene is often used in the pseudovirus system.…”

Section: Introductionmentioning

confidence: 99%

A Novel and Secure Pseudovirus Reporter System Based Assay for Neutralizing and Enhancing Antibody Assay Against Marburg Virus

Wang

Pei

et al. 2022

Front. Microbiol.

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Marburg virus (MARV) is one of the principal members of the filovirus family, which can cause fatal hemorrhagic fever in humans. There are currently no prophylactic and therapeutic drugs on the market, and the high pathogenicity and infectivity of MARV make its research highly dependent on biosafety level 4 conditions, severely hindering the development of vaccines and therapies. Therefore, the development of medicines, such as MARV serological diagnosis, vaccines, and therapeutic antibody drugs, urgently needs a safe, convenient, and biosafety level 2 detection method to measure the neutralizing activity of MARV antibodies. To this end, we report a neutralization assay relying on a Rabies virus (RABV) reverse genetic operating system. We constructed infectious clones carrying the eGFP reporter gene and the full length of the original unmodified MARV GP gene. Based on the critical parameters of phylogenetic analysis, recombinant viruses targeting representative strains in the two major MARV lineages were successfully rescued. These pseudoviruses are safe in mice, and their inability to infect cells after being neutralized by antibodies can be visualized under a fluorescence microscope. We tested the system using the neutralizing antibody MR191. MR191 can significantly block the infection of BSR cells with pseudovirus. We compared it with the traditional lentivirus-type pseudovirus system to verify the system’s credibility and obtained the same results as reported in the literature. In general, we have established a safe and visualized method for evaluating the neutralizing activity of MARV antibodies. Compared with traditional methods, it has the advantages of convenient operation, short cycle, and low cost. It is a candidate method that can replace actual viruses for a neutralization assay.

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Section: Introductionmentioning

confidence: 99%

A Novel and Secure Pseudovirus Reporter System Based Assay for Neutralizing and Enhancing Antibody Assay Against Marburg Virus

Wang

Pei

et al. 2022

Front. Microbiol.

View full text Add to dashboard Cite

show abstract

“…The neutralizing antibody detection method based on pseudovirus primarily uses HIV, VSV, and other vectors to package viral. From the perspective of biosafety, most of the packaging vectors used presently produce replication-deficient recombinant virus; that is, the virus particles obtained after packaging cannot be amplified in vitro and need to be "realtime rescue "brings experimental and economic pressure to relevant research, (Whitt, 2010;Vial et al, 2020;Salazar-Garcia et al, 2021). At the same time, a luciferase reporter gene is often used in the pseudovirus system.…”

Section: Introductionmentioning

confidence: 99%

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“…The neutralizing antibody detection method based on pseudovirus primarily uses HIV, VSV, and other vectors to package viral. From the perspective of biosafety, most of the packaging vectors used presently produce replication-deficient recombinant virus; that is, the virus particles obtained after packaging cannot be amplified in vitro and need to be "realtime rescue "brings experimental and economic pressure to relevant research, (Whitt, 2010;Vial et al, 2020;Salazar-Garcia et al, 2021). At the same time, a luciferase reporter gene is often used in the pseudovirus system.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of VSV Pseudovirus and Focus Reduction Neutralization Assays for Measurement of Anti-Andes orthohantavirus Neutralizing Antibodies in Patient Samples

Cited by 5 publications

References 17 publications

A Novel and Secure Pseudovirus Reporter System Based Assay for Neutralizing and Enhancing Antibody Assay Against Marburg Virus

A Novel and Secure Pseudovirus Reporter System Based Assay for Neutralizing and Enhancing Antibody Assay Against Marburg Virus

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