ArticleVirology journal2013
A neutralization assay for respiratory syncytial virus using a quantitative PCR-based endpoint assessment.
Article in Virology journal, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 26 citations in OpenAlex.
- Improved RSV Neutralization Assay Using Recombinant RSV Expressing Reporter Fluorescent Protein.Methods and protocols · 2025Article
- Stability of Respiratory Syncytial Virus in Nasal Aspirate From Patients Infected With RSV.Influenza and other respiratory viruses · 2024Article
- Reversible disruption of XPO1-mediated nuclear export inhibits respiratory syncytial virus (RSV) replication.Scientific reports · 2021Article
- Immunotherapeutic Efficacy of IgY Antibodies Targeting the Full-Length Spike Protein in an Animal Model of Middle East Respiratory Syndrome Coronavirus Infection.Pharmaceuticals (Basel, Switzerland) · 2021Article
- Bivalent Junin & Machupo experimental vaccine based on alphavirus RNA replicon vector.Vaccine · 2020Article
- Multiplex PCR-Based Neutralization (MPBN) Assay for Titers Determination of the Three Types of Anti-Poliovirus Neutralizing-Antibodies.Vaccines · 2020Article
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- Development of a High-Throughput Respiratory Syncytial Virus Fluorescent Focus-Based Microneutralization Assay.Clinical and vaccine immunology : CVI · 2017Article
- Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment.Journal of clinical microbiology · 2017Article
- A simple, inexpensive method for preparing cell lysates suitable for downstream reverse transcription quantitative PCR.Scientific reports · 2014Article
- Development of a luciferase immunoprecipitation system assay to detect IgG antibodies against human respiratory syncytial virus nucleoprotein.Clinical and vaccine immunology : CVI · 2014Article
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundFew studies have used quantitative polymerase chain reaction (qPCR) as an approach to measure virus neutralization assay endpoints. Its lack of use may not be surprising considering that sample nucleic acid extraction and purification can be expensive, labor-intensive, and rate-limiting.
methodsVirus/antibody mixtures were incubated for one hour at 37°C and then transferred to Vero cell monolayers in a 96-well plate format. At 24 (or 48) hours post-infection, we used a commercially available reagent to prepare cell lysates amenable to direct analysis by one-step SYBR Green quantitative reverse transcription PCR using primers specific for the RSV-N gene, thereby obviating the need for cumbersome RNA extraction and purification. The neutralization titer was defined as the reciprocal of the highest dilution needed to inhibit the PCR signal by 90% when compared with the mean value observed in virus control wells in the absence of neutralizing antibodies.
resultsWe have developed a qPCR-based neutralization assay for human respiratory syncytial virus. Due to the sensitivity of qPCR in detecting virus replication, endpoints may be assessed as early as 24 hours post-infection. In addition, the dynamic range of qPCR provides a basis for the assay to be relatively robust to perturbations in input virus dose (i.e., the assay is in compliance with the Percentage Law).
conclusionsThis qPCR-based neutralization assay is suitable for automated high-throughput applications. In addition, our experimental approach may be generalizable for the rapid development of neutralization assays for other virus families.
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