Orthogonal SARS-CoV-2 Serological Assays Enable Surveillance of Low-Prevalence Communities and Reveal Durable Humoral Immunity

Article activity feed

1. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.08.14.20174490: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Freezing and thawing had no effect on levels of antibodies detected by ELISA or PRNT.	PRNT suggested: None
   Antigens and Antiviral antibody assay: The bacterially-produced recombinant receptor-binding domain (RBD) of the spike (S) glycoprotein was a gift of Dr Daved Fremont (Washington University, St. Louis, MO)	Antigens suggested: None Antiviral suggested: None
   Plates were then washed and incubated for 1hr in 1% PBS and milk containing an anti-human IgG-HRP …

   SciScore for 10.1101/2020.08.14.20174490: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Freezing and thawing had no effect on levels of antibodies detected by ELISA or PRNT.	PRNT suggested: None
   Antigens and Antiviral antibody assay: The bacterially-produced recombinant receptor-binding domain (RBD) of the spike (S) glycoprotein was a gift of Dr Daved Fremont (Washington University, St. Louis, MO)	Antigens suggested: None Antiviral suggested: None
   Plates were then washed and incubated for 1hr in 1% PBS and milk containing an anti-human IgG-HRP conjugated antibody (Jackson Immunoresearch catalog 709-035-149) at a concentration of 1:2000 for 1 hour.	anti-human IgG-HRP suggested: None
   For IgM detection an anti-human IgM-HRP conjugated antibody (Jackson Immunoresearch catalog 709-035-073) was used at a concentration of 1:5000 and incubated for 1 hour.	an anti-human IgM-HRP suggested: None anti-human IgM-HRP suggested: (SouthernBiotech Cat# 2020-05, RRID:AB_2795603)
   A 384 well format was applied for high throughput screening, with protocol conditions remaining identical except for the substitution of anti-human Pan-Ig HRP conjugated antibody (Jackson Immunoresearch catalog 109-035-064).	anti-human Pan-Ig HRP suggested: None
   Every plate contained at least 32 seronegative controls and either CR3022 or HM3128 (Creative Diagnostics) monoclonal antibodies as a positive control for RBD or S2, respectively.	CR3022 suggested: (Imported from the IEDB Cat# CR3022, RRID:AB_2848080) S2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Briefly, Vero cells (ATCC # CCL-81) were plated in 96 well tissue culture plates and grown overnight.	Vero suggested: ATCC Cat# CCL-81, RRID:CVCL_0059)

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   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   Limitations of current study: The above assay allowed us to examine the influence of age, sex, and disease severity on levels of humoral immunity in our tested populations. Similar to other studies (Qu et al., 2020; To et al., 2020), we found that severe disease correlated positively with levels of antibody immunity. While both older adults (>50 and even more >65 years of age) and males are more vulnerable to COVID-19 (Klein et al., 2020a, 2020b; Palaiodimos et al., 2020), levels of humoral immunity did not reveal age or sex-related differences that could explain such vulnerability. A caveat here is that our study had a limited longitudinal component and that we could not determine whether there may have been a delay or reduction in humoral immunity at earlier time points of the disease. A second related caveat is that in our community testing cohort we may have missed individuals who were seropositive initially but then seroreverted by the time of the antibody test. Finally, the latest timepoint post-disease onset in our study is 122 days. It remains possible that antibody titers will wane substantially at later times. Additional serial sampling of PCR-confirmed mild cases will be required to test these possibilities.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
   • No protocol registration statement was detected.

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   Read the original source
2. Version published to 10.1016/j.immuni.2020.10.004

   Nov 1, 2020
3. 

   ScreenIT

   Aug 16, 2020

   SciScore for 10.1101/2020.08.14.20174490: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement

   not detected.

   Randomization

   not detected.

   Blinding

   not detected.

   Power Analysis

   not detected.

   Sex as a biological variable

   Figure S3 related to Figure 3 B 4 3 2 1 neg avg 10 20 30 40 days post-PCR+ neg avg 50 0 0 D 20 30 40 days post-PCR+ 20 30 40 days post-PCR+ Female Male p= 0.34 3 2 1 Neg Avg 0 10 50 4 RBD AUC (IgG) neg avg 10 0 50 20 40 60 80 100 F 4 p= 0.67 Male N AUC (IgG) Female Female Male p= 0.82 3 2 1 Neg Avg Neg Avg 0 20 40 60 80 100 0 40 60 Female Male p= 0.20 20 80 100 Figure S3 related to Figure 3: Antibody responses to SARS-Cov2 in asymptomatic individuals and in females and males.

   Cell Line Authentication

   not detected.

   Table 2: Resources

   Antibodies
   Sentences	R…

   SciScore for 10.1101/2020.08.14.20174490: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement

   not detected.

   Randomization

   not detected.

   Blinding

   not detected.

   Power Analysis

   not detected.

   Sex as a biological variable

   Figure S3 related to Figure 3 B 4 3 2 1 neg avg 10 20 30 40 days post-PCR+ neg avg 50 0 0 D 20 30 40 days post-PCR+ 20 30 40 days post-PCR+ Female Male p= 0.34 3 2 1 Neg Avg 0 10 50 4 RBD AUC (IgG) neg avg 10 0 50 20 40 60 80 100 F 4 p= 0.67 Male N AUC (IgG) Female Female Male p= 0.82 3 2 1 Neg Avg Neg Avg 0 20 40 60 80 100 0 40 60 Female Male p= 0.20 20 80 100 Figure S3 related to Figure 3: Antibody responses to SARS-Cov2 in asymptomatic individuals and in females and males.

   Cell Line Authentication

   not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   ( C) Neutralizing titers correlate with IgM antibody titers for RBD.	IgM suggested: None
   Figure S2 related to Figure 2: Optimization of secondary screens for S2 and N. (A) N-reactive IgM antibody titers were correlated to PRNT90 neutralization titers.	N-reactive IgM suggested: None
   Supernatant and cell lysate were combined, subjected to a single freeze-thaw, and then centrifuged at 3000RPM Antigens and Antiviral antibody assay: The bacterially-produced recombinant receptor-binding domain (RBD) of the spike (S) glycoprotein was a gift of Dr Daved Fremont (Washington University, St. Louis, MO)	Antiviral suggested: None
   Plates were then washed and incubated for 1hr in 1% PBS and milk containing an anti-human IgG-HRP conjugated antibody (Jackson Immunoresearch catalog 709-035-149) at a concentration of 1:2000 for 1 hour.	anti-human IgG-HRP suggested: None
   For IgM detection an anti-human IgM-HRP conjugated antibody (Jackson Immunoresearch catalog 709035-073) was used at a concentration of 1:5000 and incubated for 1 hour.	an anti-human IgM-HRP suggested: None anti-human IgM-HRP suggested: (SouthernBiotech Cat# 2020-05, RRID:AB_2795603)
   A 384 well format was applied for high throughput screening, with protocol conditions remaining identical except for the substitution of anti-human Pan-Ig HRP conjugated antibody (Jackson Immunoresearch catalog 109-035-064).	anti-human Pan-Ig HRP suggested: None
   Every plate contained at least 32 seronegative controls and either CR3022 or HM3128 (Creative Diagnostics) monoclonal antibodies as a positive control for RBD or S2, respectively.	CR3022 suggested: (Imported from the IEDB Cat# CR3022, RRID:AB_2848080) S2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Coronavirus 2, Isolate USA-WA1/2020 (BEI NR-52281) was passaged once on Vero (ATCC #CCL-81) cells at a MOI of 0.01 for 72 hours.	Vero suggested: ATCC Cat# CCL-81, RRID:CVCL_0059)

   ---

   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

   ---

   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   Limitations of current study: The above assay allowed us to examine the influence of age, sex, and disease severity on levels of humoral immunity in our tested populations. Similar to other studies (Qu et al., 2020; To et al., 2020), we found that severe disease correlated positively with levels of antibody immunity. While both older adults (>50 and even more >65 years of age) and males are more vulnerable to COVID-19 (Klein et al., 2020a, 2020b; Palaiodimos et al., 2020), levels of humoral immunity did not reveal age or sex-related differences that could explain such vulnerability. A caveat here is that our study had a limited longitudinal component and that we could not determine whether there may have been a delay or reduction in humoral immunity at earlier time points of the disease. A second related caveat is that in our community testing cohort we may have missed individuals who were seropositive initially but then seroreverted by the time of the antibody test. Finally, the latest timepoint postdisease onset in our study is 122 days. It remains possible that antibody titers will wane substantially at later times. Additional serial sampling of PCR-confirmed mild cases will be required to test these possibilities.

   ---

   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

   ---

   Results from JetFighter: We did not find any issues relating to colormaps.

   ---

   About SciScore

   SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

   Read the original source
4. 

   Version published to 10.1101/2020.08.14.20174490 on medRxiv

   Aug 16, 2020