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Sars cov 2 svnt kit

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The SARS-CoV-2 sVNT Kit is a laboratory-based serological test designed to detect the presence of neutralizing antibodies against the SARS-CoV-2 virus. The kit utilizes a surrogate virus neutralization test (sVNT) method to provide an indication of an individual's neutralizing antibody levels.

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Lab products found in correlation

Liaison sars cov 2 s1 s2 igg kit, by DiaSorin (2 mentions) Eti max 3000, by DiaSorin (2 mentions) Varioskan lux, by Thermo Fisher Scientific (1 mentions) Svnt assay, by GenScript (1 mentions) Elecsys anti sars cov 2, by Roche (1 mentions) Sars cov 2 igg, by Abbott (1 mentions) Quantivac, by EUROIMMUN (1 mentions) Cpass sars cov 2 neutralization antibody detection kit, by GenScript (1 mentions) Thermo varioskan flash, by Thermo Fisher Scientific (1 mentions) Eti max 3000 equipment, by DiaSorin (1 mentions)

22 protocols using sars cov 2 svnt kit

1

Neutralizing Antibody Dynamics Against SARS-CoV-2

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At the same time, we evaluated the anti-RBD responses in fasting blood samples at three-time points above by the serum surrogate virus neutralization test (sVNT) to assess the dynamic changes of the neutralizing antibody. Circulating NAb against SARS-CoV-2 was detected which blocked the interaction between the RBD of the viral spike glycoprotein with the angiotensin-converting enzyme 2 cell surface receptor in the experiment. Recombinant S-RBD from the wild type (Wuhan-Hu-1) and omicron (B.1.1.529) strains were used in this study. All experimental operations and the use of the SARS-CoV-2 sVNT kit (GenScript) were performed according to the manufacturer's instructions. Tests were performed on Varioskan Lux (ThermoFisher).
For all assays, the limit of quantitation (LOQ) was 9.38 U/mL, and levels < LOQ were substituted with LOQ/Sqr(2).
Deng Y., Huang L., Liu P., Geng X., Lin Z., Zheng Z., Zhan M., Zhang Z., Liu J, & Sun T. (2023). Association of fat-soluble vitamins (A, D, and E) status with humoral immune response to COVID-19 inactivated vaccination. Frontiers in Nutrition, 10, 1167920.
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2

Serology Assessment of COVID-19 Vaccine Response

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Serology was assessed for all participants prior to the first vaccine dose, at D69 (interval from second dose to serum collection = 6 weeks) and 6 months after the second dose (D210). The blood samples were collected, centrifuged, and stored at −80 °C until analysis. The serologic assay consisted of the measurement of the total IgG antibodies against the SARS-CoV-2 S1 and S2 proteins performed by chemiluminescent immunoassay on the ETI-MAX-3000 equipment (DiaSorin, Italy) using the Indirect ELISA, LIAISON® SARS-CoV-2 S1/S2 IgG kit Cat# 311450 and Cat# 311451 (DiaSorin, Italy), and the measurement of the circulating NAb against SARS-CoV-2 using the SARS-CoV-2 sVNT Kit, Cat# L00847-A (GenScript, Piscataway, NJ, USA). The two assays were performed following the manufacturer’s instructions. Samples with 15.0 UA/mL or more for total anti-S1/S2 IgG and with 30% or more inhibition for neutralizing assay, were considered seropositive according to the manufacturer’s guide29 ,30 (link). Furthermore, quantitative results were reported, attributing the value of 1.9 UA/mL (half of the lower limit of quantification 3.8 UA/mL) to undetectable levels (<3.8 UA/mL) of total IgG and 15% (half of the lower limit of quantification 30%) to undetectable levels (<30%) of neutralizing antibodies.
Silva C.A., Medeiros-Ribeiro A.C., Kupa L.V., Yuki E.F., Pasoto S.G., Saad C.G., Fusco S.R., Pereira R.M., Shinjo S.K., Halpern A.S., Borba E.F., Souza F.H., Guedes L.K., Miossi R., Bonfiglioli K.R., Domiciano D.S., Shimabuco A.Y., Andrade D.C., Seguro L.P., Fuller R., Sampaio-Barros P.D., Assad A.P., Moraes J.C., Goldenstein-Schainberg C., Giardini H.A., Silva H.C., Martins V.A., Villamarin L.E., Novellino R.S., Sales L.P., Araújo C.S., Silva M.S., Filho D.M., Lopes M.H., Duarte A.J., Kallas E.G., Aikawa N.E, & Bonfa E. (2022). Immunogenicity decay and case incidence six months post Sinovac-CoronaVac vaccine in autoimmune rheumatic diseases patients. Nature Communications, 13, 5801.
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3

SARS-CoV-2 Neutralization Assay

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The spike protein (S) receptor binding domain (RBD) is responsible for recognizing the cell surface receptor, angiotensin converting enzyme-2 (ACE2). The RBD of SARS-CoV-2 S protein strongly interacts with hACE2. The SARS-CoV-2 sVNT Kit (Genscript; Leiden, Netherlands) is a blocking ELISA, which mimics this virus receptor binding process. The protein-protein interaction between a horseradish peroxidase (HRP) conjugated recombinant SARS-CoV-2 RBD fragment (HRP-RBD) and hACE2 can be blocked by neutralizing antibodies against the SARS-CoV-2 RBD and residual HRP activity is measured as a surrogate for neutralization. The assay was performed with 1:50 diluted plasma following the manufacturer’s instructions.
von Rhein C., Scholz T., Henss L., Kronstein-Wiedemann R., Schwarz T., Rodionov R.N., Corman V.M., Tonn T, & Schnierle B.S. (2020). Comparison of potency assays to assess SARS-CoV-2 neutralizing antibody capacity in COVID-19 convalescent plasma. Journal of Virological Methods, 288, 114031.
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4

SARS-CoV-2 sVNT Antibody Neutralization Assay

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A SARS-CoV-2-specific surrogate Virus Neutralization Test (sVNT) was used to study the levels of plasma neutralizing antibodies against SARS-CoV-2 S-RBD (SARS-Cov-2 sVNT kit, GenScript, USA, Inc.) according to the manufacturer's protocol. Evaluation of the data was performed by following the manufacturer's recommendations for positive and negative cutoffs. The results were obtained by calculating inhibition rates for samples per the following equation: Inhibition = (1 – sample O.D./O.D. negative control) × 100%. Neutralizing antibody levels >20% were considered to be positive.
Ali H., Alahmad B., Al-Shammari A.A., Alterki A., Hammad M., Cherian P., Alkhairi I., Sindhu S., Thanaraj T.A., Mohammad A., Alghanim G., Deverajan S., Ahmad R., El-Shazly S., Dashti A.A., Shehab M., Al-Sabah S., Alkandari A., Abubaker J., Abu-Farha M, & Al-Mulla F. (2021). Previous COVID-19 Infection and Antibody Levels After Vaccination. Frontiers in Public Health, 9, 778243.
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5

SARS-CoV-2 Spike Protein Antibody Detection

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A SARS-CoV-2 sVNT kit (GenScript) was used to detect antibodies that block the interaction between the SARS-CoV-2 spike protein and ACE2, as described previously (16 (link)). HRP-conjugated recombinant SARS-CoV-2 RBD fragment bound to neutralizing antibodies, preventing capture by the human ACE2 protein in the well, which was subsequently removed in the following wash step. Substrate was added and incubated for 20 minutes at room temperature and results measured by spectrophotometry. Color intensity was inversely dependent on the titer of anti–SARS-CoV-2 neutralizing antibodies.
Habel J.R., Chua B.Y., Kedzierski L., Selva K.J., Damelang T., Haycroft E.R., Nguyen T.H., Koay H.F., Nicholson S., McQuilten H.A., Jia X., Allen L.F., Hensen L., Zhang W., van de Sandt C.E., Neil J.A., Pragastis K., Lau J.S., Jumarang J., Allen E.K., Amanant F., Krammer F., Wragg K.M., Juno J.A., Wheatley A.K., Tan H.X., Pell G., Walker S., Audsley J., Reynaldi A., Thevarajan I., Denholm J.T., Subbarao K., Davenport M.P., Hogarth P.M., Godfrey D.I., Cheng A.C., Tong S.Y., Bond K., Williamson D.A., McMahon J.H., Thomas P.G., Pannaraj P.S., James F., Holmes N.E., Smibert O.C., Trubiano J.A., Gordon C.L., Chung A.W., Whitehead C.L., Kent S.J., Lappas M., Rowntree L.C, & Kedzierska K. (2023). Immune profiling of SARS-CoV-2 infection during pregnancy reveals NK cell and γδ T cell perturbations. JCI Insight, 8(7), e167157.
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6

Quantification of SARS-CoV-2 Antibodies

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Enzyme-linked immunosorbent assay (ELISA) kit (SERION ELISA agile SARS-CoV-2 IgG and IgA SERION Diagnostics, Würzburg, Germany) was used to measure plasma levels of SARS-CoV-2-specific IgG antibodies based on the manufacturer's protocol. Units of IgG levels were reported as binding antibody units (BAU)/ml. Values below 31.5 BAU/ml were considered negative or non-protective.
The SARS-CoV-2-Specific Neutralizing Antibodies levels in the plasma were quantified using a SARS-CoV-2-specific surrogate Virus Neutralization Test (sVNT) (SARS-CoV-2 sVNT kit, GenScript, USA, Inc.) based on the manufacturer's protocol. Determination of positive and negative thresholds was performed by following the manufacturer's recommendations. The results were interpreted by calculating inhibition rates for samples per the following equation: Inhibition = (1–sample O.D/O.D negative control) ×100%. Neutralizing antibody levels below 20% were considered negative or non-protective.
Shehab M., Alrashed F., Alfadhli A., Alsayegh A., Aldallal U., Alsayegh M., Cherian P., Alkhair I., Thanaraj T.A., Channanath A., Dashti A.A., Albanaw A., Ali H., Abu-Farha M., Abubaker J, & Al-Mulla F. (2022). Immunogenicity of BNT162b2 Vaccine Booster Dose in Patients With Inflammatory Bowel Disease Receiving Infliximab Combination Therapy: A Prospective Observational Study. Frontiers in Medicine, 9, 933996.
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7

SARS-CoV-2 sVNT Assay for Neutralization

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The SARS-CoV-2 sVNT Kit (Genscript, Leiden, Netherlands) is a blocking ELISA that mimics the virus receptor-binding process. Inhibition of the protein–protein interaction between a horseradish peroxidase (HRP)-conjugated recombinant SARS-CoV-2 spike receptor-binding domain fragment (HRP-RBD) and hACE2 is measured as a surrogate for neutralization. The assay was performed with different concentrations of EGCG or EC following the manufacturer’s instructions. The negative (non-inhibiting) and positive (inhibiting) controls of the sVNT kit were included. Inhibition was calculated following the manufacturer’s protocol using the following equation: Inhibition in % = (1 – OD of sample/OD of negative control)×100. The cutoff was set to 0, the value of the negative control.
Henss L., Auste A., Schürmann C., Schmidt C., von Rhein C., Mühlebach M.D, & Schnierle B.S. (2021). The green tea catechin epigallocatechin gallate inhibits SARS-CoV-2 infection. The Journal of General Virology, 102(4), 001574.
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8

Detecting Neutralizing Antibodies against SARS-CoV-2

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Antibodies that block the interaction between RBD and ACE2 were detected by the SARS-CoV-2 sVNT Kit (GenScript). Briefly, the serum samples and controls were pre-incubated with RBD-HRP to allow binding between antibodies and RBD-HRP. Then, the mixture was added to a capture plate pre-coated with the hACE2 protein. The unbound RBD-HRP and any RBD-HRP bound to non-neutralizing antibodies will be captured on the plate, while the NAbs/RBD-HRP complexes remain in the supernatant and are removed during washing. After washing steps, TMB solution is added. After adding the stop solution, this final solution was read at 450 nm in a microtiter plate reader. The absorbance of the sample was inversely dependent on the titer of the RBD-ACE2 blocking antibodies. According to the sVNT Kit, the cutoff value was set to 30%.
Long Y., Sun J., Song T.Z., Liu T., Tang F., Zhang X., Ding L., Miao Y., Zhu W., Pan X., An Q., Qin M., Tong X., Peng X., Yu P., Zhu P., Xu J., Zhang X., Zhang Y., Liu D., Chen B., Chen H., Zhang L., Xiao G., Zuo J., Tang W., Zhou J., Li H., Xu Z., Zheng H.Y., Long X.Y., Qin Q., Gan Y., Ren J., Huang W., Zheng Y.T., Jin G, & Gong L. (2022). CoVac501, a self-adjuvanting peptide vaccine conjugated with TLR7 agonists, against SARS-CoV-2 induces protective immunity. Cell Discovery, 8, 9.
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9

SARS-CoV-2 Neutralizing Antibody Assay

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SARS-CoV-2 sVNT Kit (cPAss from Genscript) was used to evaluate the neutralizing capacity of anti-SARS-CoV-2 antibodies present in the serum. This is a blocking enzyme-linked immunosorbent assay (ELISA), which mimics the virus-host interaction. Binding of a horseradish peroxidase conjugated RBD-fragment of the SARS-CoV-2 (HRP-RBD) to the human host ACE2 receptor can be blocked by neutralizing antibodies against the SARS-CoV-2 spike protein, containing the RBD in the serum or plasma. The strength of HRP signal indicates the degree of blockage and therefore indirect the neutralizing capacity. The sVNT assay from Genscript has been validated and described previously (7 (link)–10 (link)).
Chu C., Schönbrunn A., Klemm K., von Baehr V., Krämer B.K., Elitok S, & Hocher B. (2022). Impact of hypertension on long-term humoral and cellular response to SARS-CoV-2 infection. Frontiers in Immunology, 13, 915001.
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10

Evaluating SARS-CoV-2 Antibody Assays

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Seven serological assays were assessed in this study: three CLIAs [SARS-CoV-2 IgG (Abbott, Chicago, IL, United States); Elecsys Anti-SARS-CoV-2 (Roche, Basel, Switzerland); ADVIA Centaur SARS-CoV-2 Total (Siemens, Munich, Germany)], three LFIAs [STANDARD F COVID-19 IgM/IgG Combo FIA (SD Biosensor Inc., Gyeonggi-do, Korea), briefly SDF; STANDARD Q COVID-19 IgM/IgG Combo (SD Biosensor Inc.), briefly SDQ; P4DETECT COVID-19 IgG/IgM (PRIME4DIA Co., Gyeonggi-do, Korea), briefly P4D], and one SARS-CoV-2 sVNT kit (GenScript Biotech Co., NJ, United States) (Table 1). All samples were analyzed using six SARS-CoV-2 bAb assays. Because of insufficient sample volumes, only 418 serum samples, consisting of 385 samples from COVID-19-positive patients and 33 samples from COVID-19-negative patients with false-positive results from at least one of the binding antibody assays, were subjected to the SARS-CoV-2 sVNT testing. All assays were performed at the Diagnostic Immunology Laboratory of CNUH according to the manufacturer’s instructions.
Choi H.W., Jeon C.H., Won E.J., Kang S.J., Lee S.Y, & Kee S.J. (2022). Performance of Severe Acute Respiratory Syndrome Coronavirus 2 Serological Diagnostic Tests and Antibody Kinetics in Coronavirus Disease 2019 Patients. Frontiers in Microbiology, 13, 881038.
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