Plaque Reduction Neutralization Testing (PRNT) to Assess Antiviral Activity

The Plaque Reduction Neutralization Test (PRNT), often referred to as a PRNT assay, is used to measure the ability of antibodies or serum to neutralize a virus. It is a highly specific and quantitative method for determining the presence and potency of neutralizing antibodies in serum or other biological samples. The PRNT assay is commonly employed in virology research, vaccine development, and seroepidemiology. 

Why Choose IBT for Plaque Reduction Neutralization Testing?

IBT Bioservices is a leader, in providing in vitro services such as the PRNT assay for researchers and organizations focusing on vaccine & therapeutic development, antiviral treatments, and serological studies. Our extensive experience with plaque reduction neutralization assays ensures you receive accurate, reproducible data, with results validated through stringent quality control measures. We use the latest technologies to provide tailored solutions that meet your specific research requirements, making us the go-to partner for PRNT assays.

Our Process

Our plaque reduction neutralization testing process is designed to deliver reliable and precise results:

Sample Preparation: Test articles or serum samples are prepared and diluted.

Virus-Antibody Incubation: The virus is mixed with diluted serum or test articles and incubated with host cells.

Methylcellulose Overlay: After a set incubation period, the cells are overlaid with methylcellulose, which restricts virus spread.

Plaque Counting: Plaques are visualized using crystal violet staining, and the number of plaques is counted to assess neutralizing activity.

PRNT50 Calculation: The serum dilution at which 50% of the virus is neutralized is determined as the PRNT50 value.

 

Applications & Features of PRNT

  • Vaccine & Therapeutic Development: The PRNT assay is instrumental in assessing the ability of vaccines to induce neutralizing antibodies that protect against viral infections.
  • Virology Research: PRNT assays are widely used to study the immune response to viral infections, evaluate antiviral therapies, and understand the mechanisms of viral neutralization.

Epidemiology: PRNT assays are essential in seroepidemiological studies, helping to determine the prevalence of specific antibodies within populations and the level of exposure to particular viruses.

Our Process

We offer a large panel of viral strains for PRNT testing, including:

  • Dengue virus (Serotypes 1-4)
  • Zika virus
  • Influenza A (H1N1, H3N2)
  • Respiratory Syncytial Virus (RSV)
  • Herpes Simplex Virus
  • Chikungunya Virus

In addition, we can develop custom PRNT assays for emerging pathogens or specific research requirements.

Small molecule inhibitors are tested against Dengue virus serotype 2 (DENV-2) Y98P in a PRNT assay using Vero cells

Test articles (TAs) 1-4 are small molecule inhibitors tested against

DENV-2 Y98P in Vero cells in a PRNT assay. Dilutions of test articles were incubated with a known amount of virus and both test article dilutions and virus were added to cells for a set period of time before the addition of a methylcellulose layer. After a predetermined amount of time, plaques were counted and compared to the virus only control to calculate PRNT50 values. 

 

All TAs show neutralizing activity against DENV-2 Y98P with TA2 exhibiting the most neutralization of D2 Y98P. The IBT internal control is a mouse monoclonal antibody against the Dengue envelope protein.

Zika virus (ZIKV) antiserum was tested for ZIKV neutralization in a PRNT assay

Antiserum was tested in a PRNT assay against ZIKV FSS13025. The antiserum was serially diluted for 12 dilutions and was incubated with ZIKV FSS13025 at a predetermined MOI. The antiserum and virus mixture was added to Vero cells in duplicate and a methylcellulose layer was later added. The cells were incubated for several days and then stained with crystal violet to visualize the ZIKV FSS13025 plaques. The lack of plaques in the first dilution indicates that the control is neutralizing toward ZIKV as no infection is observed.

The PRNT assay is based on the principle that neutralizing antibodies, present in a test serum, have the capacity to neutralize the infectivity of a virus by preventing it from infecting host cells. The test measures the dilution of serum at which 50% of the virus is neutralized, known as the 50% neutralization titer (PRNT50).

OUR PATHOGENS

We have a large panel of strains available and the ability to develop the assay for previous and emerging influenza strains.

We offer an extensive range of viruses for your testing needs and can develop custom PRNT assays to meet your specific research objectives.

Respiratory Viruses – Influenza, RSV, ADV

Arboviruses – Dengue 1-4, CHKV, JEV, LACV, RVF, YFV, ZIKV

Herpes Viruses – HSV-1, HSV-2, CMV

Enteric Viruses – Enteroviruses, Echoviruses, CVA

Pseudoviruses – Filoviruses, SARS-CoV-2

With years of experience in the field, our team of skilled scientists and technicians are well-versed in conducting PRNT assays for a wide range of pathogens, including influenza viruses, dengue viruses, Respiratory Syncytial Virus (RSV), and Herpes Simplex Virus (Plaque Reduction Neutralization Test ). We utilize state-of-the-art equipment and follow stringent quality control measures to ensure precise and reproducible results.

OUR EXPERTISE

Key applications of the PRNT Assay

Vaccine Development

The PRNT assay is instrumental in assessing the ability of vaccines to induce neutralizing antibodies in individuals.

Virology Research

It is widely used in the study of the immune response to viral infections, the evaluation of antiviral treatments, and understanding the mechanisms of viral neutralization.

Epidemiology

PRNT assays are utilized in seroepidemiological studies to determine the prevalence of specific antibodies within populations, helping to estimate the extent of exposure to particular viruses.

Disease Diagnosis

Some PRNT assays are used for diagnostic purposes to confirm the presence of neutralizing antibodies to specific viruses, especially in the context of certain viral diseases.

WHY CHOOSE IBT?

EXPERTISE

Our team of scientists and technicians are highly skilled and experienced in conducting HAI assays, ensuring accurate and reliable results.

CUSTOMIZED SOLUTIONS

We understand the unique needs of each research project and provide customized assay designs to meet your specific requirements.

TIMELY RESULTS

We prioritize efficiency without compromising on quality. Our streamlined processes and advanced equipment allow for quick turnaround times, ensuring you receive results promptly.

QUALITY ASSURANCE:

We maintain rigorous quality control measures throughout our operations, ensuring the accuracy and reproducibility of our assay results.

EXCEPTIONAL SUPPORT:

Our dedicated customer support team is always available to address your queries and provide assistance at every step of the process.

FAQ

What is the plaque reduction assay method?
The plaque reduction assay is a technique used to measure the neutralizing ability of antibodies or antiviral agents. It involves incubating a virus with test serum or antiviral agents, followed by infection of host cells. After incubation, the cells are stained, and the reduction in plaque formation is measured to determine the neutralizing activity.

What is a focus reduction neutralization test?
The focus reduction neutralization test (FRNT) is similar to PRNT but is used for viruses that do not form clear plaques. Instead of plaques, viral foci (areas of infection) are counted. FRNT is commonly used for viruses like Zika or Dengue when clear plaque formation is not observed.

What is a plaque reduction neutralization test (PRNT)?

The plaque reduction neutralization test (PRNT) is used to measure the neutralizing antibody response to a specific virus. It is a key assay in vaccine development and seroepidemiology to determine the presence and potency of antibodies against various viruses, including but not limited to Dengue virus, Zika virus, and West Nile virus. The PRNT50 value indicates the serum dilution that neutralizes 50% of the viral plaques, making it an essential tool in virology research.

Contact us today at services@ibtbioservices.com to learn more about our Plaque Reduction Neutralization Testing (PRNT) services and how we can support your research in virology, vaccine & therapeutic development, or serological studies.

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