Covid-19 assays and studies

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covid-19 ASSAYS and studies

IITRI can perform the following COVID-19 assays and studies under BSL-3 using SARS-CoV-2. Currently available strains: Washington (wildtype USA-WA1/2020), UK B.1.1.7 , South African B.1.35, and Brazilian P.1.

 

Efficacy assay (Immunostain)

– Efficacy assay using either Vero E6 or Calu-3 cells

– Looks at 50% effective concentration (EC50) of the drug against SARS-CoV-2. 

SARS-CoV-2 efficacy assay with immunostain

 

Neutralization assays (PRNT, ELISA)

SARS-CoV-2 48hr neutralization assays for antibodies to determine reduction of plaques and viral measurements.

PRNT

– Plaques are visualized and counted using an ELISPOT instrument

– Plaque counts are fitted to a dose-response curve and plaque reduction neutralization test titers (PRNT) expressed in terms of conventional 50% PRNT end-point titers.

ELISA

– For each well, the inhibition of virus by neutralizing antibodies is calculated as the percentage of reduction of the absorbance value in respect of the virus control

– The neutralizing antibody titer is defined as the reciprocal dilution that caused 50% reduction of the absorbance value of the virus control (50% A450 reduction)

In vivo studies

Hamster model

– General clinical observations

– Body weight loss

– Viral load in nasal washes (TCID50 for live virus, SARS-CoV-2-specific RTqPCR)

– Tissue viral burden by TCID50 and/or RT-qPCR

– Histopathology of tissues, to be evaluated with standard H.E. staining

– Neutralizing antibody titer

– Cytokine expression by RT-qPCR

 

More about this model: IITRI COVID-19 hamster model

Transgenic mouse model

 – General clinical observations

– Body weight loss, mortality

– Tissue viral burden by TCID50 and/or RT-qPCR neutralizing antibody titer

– Histopathology of tissues, to be evaluated with standard H.E. staining

– Cytokine expression by RT-qPCR and/or ELISA

 

Dose range-finding data for the hACE2- transgenic mouse model

 

More about this model: IITRI COVID-19 mouse model

Ferret model

– Favorable model for looking at effects solely in upper respiratory tract

– Infection model only, no clinical signs

– Viral load and shedding in nasal washes

– Viral load in nasal washes (TCID50 for live virus, SARS-CoV-2-specific RTqPCR)

– Tissue viral burden by TCID50 and/or RT-qPCR

– Histopathology of tissues, to be evaluated with standard H.E. staining

– Neutralizing antibody titer

– Cytokine expression by RT-qPCR (ELISA reagents for ferrets are very limited)

Read more about our ferret models: IITRI COVID-19 ferret model, IITRI influenza ferret model

IITRI has established itself as a leading COVID-19 CRO and due to the importance for this testing during the pandemic, has made provisions to turn these assays and studies around rapidly.  Other related capabilities include a BSL-3/ABSL-3 aerobiology environment for inhaled drug challenge studies.

 

Contact us for more detailed information, to discuss with a PhD study director or request a cost estimate.