hauenstein scasm v1 · 2020. 12. 7. · s1igg 97 92.4% 85.7 –96.1 s2igg 46 43.8% 34.7 –53.4...

11/10/20

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:Contributing to the COVID-19 Battle Advanced Tools for SARS-CoV-2

Scott Hauenstein, PhDNovember 11, 2020

SCASM 407-012-20 Hauenstein

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Containing the spread and decreasing mortality of SARS-CoV-2 infection

Surveillance• Testing to gain information at the population level

rather than individual level

Screening• Looking for COVID-19 occurrence at

the individual level even if there is no reason to suspect exposure

Diagnosis• A test intended to diagnose an infection in

an individual suspected to have COVID-19 orbeen exposed

https://www.fda.gov/medical-devices/emergency-situations-medical-devices/faqs-testing-sars-cov-2SCASM 407-012-20 Hauenstein

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COVID-19 Global Cases

https://coronavirus.jhu.edu/map.html. Accessed Nov 9, 2020. SCASM 407-012-20 Hauenstein

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Detection of anti-SARS-CoV-2 Ab indicate prevalence may be muchhigher than RNA testing

CDC cross-sectional study performed serologic testing on a sample of residual sera obtained from persons of all ages from 10 sites across United States

Nature (2020). https://doi.org/10.1038/s41586-020-2912-6JAMA Intern Med. Published online July 21, 2020. doi:10.1001/jamainternmed.2020.4130

For m ost sites, it is likely that greater than 10 tim es [6-24] m ore SARS-CoV-2 infections occurred than the num ber of reported COVID-19 cases

Retrospective, repeated cross-sectional analysis of anti-SARS-CoV-2 spike antibodies in weekly intervals from Feb to July 2020 using more than 10,000 plasma samples from patients at Mount Sinai Hospital in NYC

Stable seroprevalence of ~22% in NYC


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Overview of Bio-Rad diagnostic tests

JAMA. 2020;323(22):2249-2251.doi:10.1001/jama.2020.8259SCASM 407-012-20 Hauenstein

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SARS-CoV-2 Droplet Digital PCR KitAn Em ergency Use Authorized Test for Diagnosing COVID-19

● Intended for the qualitative detection of nucleic acids from SARS-CoV-2

● Triplex assay contains primers and probes targeting specific regions unique to the SARS-CoV-2 virus with low risk of mutation

○ N1, N2, RPP30

○ NTC, positive and extraction control included

● 5-279 patient samples per day on a single platform

● LOD ~150 copies/mL


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https://www.fda.gov/medical-devices/emergency-situations-medical-devices/faqs-testing-sars-cov-2https://coronavirus.jhu.edu/map.html

11/10/20

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Wastewater Surveillance


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Advanced detection of SARS-CoV-2 before outbreak

Red curve = SARS-CoV-2Grey curve = Hospital admissions per day (left) or New COVID-19 cases per day (right)

Hospital Admissions (3 day advance peak) COVID-19 Cases (7 day advance peak)

SARS-CoV-2 RNA concentrations in primary municipal sewage sludge as a leading indicator of COVID-19 outbreak dynamics. doi: https://doi.org/10.1101/2020.05.19.20105999SCASM 407-012-20 Hauenstein

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Utility of Serological Testing• Should Use Validated Tests

• >55 FDA EUA available (as of November 2020)• Detect different antigens (spike 1, spike 2, receptor binding domain, nucleocapsid protein)

h t tp s : / /w w w .b io p h y s ic s .o r g /b lo g /c o r o n a v i r u s - s t r u c tu r e - v a c c in e - a n d - th e r a p y - d e v e lo p m e n t SCASM 407-012-20 Hauenstein

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https://www.biophysics.org/blog/coronavirus-structure-vaccine-and-therapy-development

11/10/20

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IDSA guidelines

8 major recommendations

Within recommendations, panel identified three clinical scenarios where antibody testing was

felt to have potential utility for diagnosis.

1.Serosurveillance epidemiology studies

2.Helpful in the evaluation of individual patients with a high clinical suspicion for COVID-19 when the results of molecular diagnostic testing are repeatedly negative or such testing was not performed.

Total Ab or IgG assay should be performed 3-4 weeks aftersymptom onset

3.Useful for assessments of suspected multisystem inflammatory syndrome in children.

https://www.idsociety.org/practice-guideline/covid-19-guideline-serology/ SCASM 407-012-20 Hauenstein

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Platelia SARS-CoV-2 Total Antibody Test

Immunoassay kit to screen for total antibodies (IgM, IgA, IgG) to SARS-CoV-2

● open microplate format● run manual or fully automated (Evolis)● detection in human serum or plasma

For in vitro diagnostic use by professionals in a laboratory environment only. This kit is for Emergency Use Authorization testing only.

C o n fig u ratio n T ests /K it

T o ta l - A n tibody test

P o s /N eg K it

C o n tro ls

B io -R ad S AR S -C o V -2

E U A k it

M an u al o rA utom atedIn stru m en ts

Serology Kit 5 Plates x 96 wells 480 IgA, IgM, IgG ✔ ✔ ✔


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Clinical Performance Characteristics: Specificity

A total of 600 specimens (500 from blood donors and 100 from hospitalized asymptomatic patients) collected prior to the outbreak of the COVID-19 pandemic were tested.

Overall specificity of 99.3% (95% CI of 98.3-99.8%)


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Clinical Performance Characteristics: SensitivityA longitudinal study was perform ed on 50 patients in the intensive care unit at 3 hospitals with clinical symptom s of COVID-19 and with a SARS-CoV-2 PCR positive result.

• Between 1 and 5 specimens per patient were collected from 2 to 92 days post onset of clinical symptoms.

Overall sensitivity of 98% (95% CI of 89.51-99.65%)


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“found Abbott, Affinity, and Bio-Rad to have the highest clinical sensitivity and specificitycom pared to the other EIAs”


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http://www.idsociety.org/practice-guideline/covid-19-guideline-serology/http://www.idsociety.org/practice-guideline/covid-19-guideline-serology/

11/10/20

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BioPlex 2200 SARS-CoV-2 IgG Panel*

IgG antibodies for RBD, S1, S2, Nucleocapsid m arkers-Composite result (qualitative) as well as a discriminatory resultfor target (semi-quant; U/mL)

Dynamic range 1-100 U/m L-onboard dilutions 1:8, 1:16, 1:32

Kit size: 200 tests per pack, 5uL sam ple

Calibrator: 5 calibrator levels (in 6 vials) per set (4PL)

Control: 2 levels of control (in 3 vials, 2 per box)

First result 45 m ins, 1 new result every 36 secs

*Pending EUA Approval SCASM 407-012-20 Hauenstein

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Sensitivity and Specificity

Cohort: Pre-COVID-19 samples (apparentlyhealthy donors)

Marker Total Negative Specificity (%)

CoV-2 IgG 1132 1128 99.6%RBD IgG 1132 1131 99.9%S1 IgG 1132 1131 99.9%S2 IgG 1132 1131 99.9%Nucleocapsid 1132 1130 99.8%

Preliminary data

Cohort: COVID-19 PCR positive samples>15 days after onset of symptoms

Marker Total % Positive 95% CICoV-2 IgG 101 96.2% 90.6 – 98.5RBD IgG 99 94.3% 88.1 – 97.4S1 IgG 97 92.4% 85.7 – 96.1S2 IgG 46 43.8% 34.7 – 53.4Nucleocapsid 98 93.3% 86.9 – 96.7


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BioPlex 2200 Summary


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Thank you!

Questions?

[email protected]


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mailto:[email protected]

hauenstein scasm v1 · 2020. 12. 7. · s1igg 97 92.4% 85.7 –96.1 s2igg 46 43.8% 34.7 –53.4...

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