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Authors: Pape and Remme et al. 

Journal/ Pre-Print:bioRxiv 

TagsMicroscopy, image analysis 

Research Highlights

  1. Development of a semi-automated imaging pipeline for SARS-CoV2 antibody detection that can be tuned for desired specificity/sensitivity 

  1. Development of a Fiji based open-source software- The PlateViewer for interactive visualization of high-throughput microscopy data- images, segmentation and resulting plots. 

Summary

The researchers developed a semi-quantitative immunofluorescence-based assay for detection of Sars-CoV-2 specific antibodies in human serum with higher specificity and sensitivity than ELISASARS-CoV2 infected VeroE6 are incubated with patient or negative control sera (for background correction) and anti-dsRNA antibody followed by labelled anti-human IgG, A and MAcquired images are pre-processed to remove barrel artefacts and cells are segmented via seeded watershed; dsRNA staining is used to classify each cell as infected or not infected. The scoring of background corrected serum channels (IgG, A and M) is computed as ratio of median fluorescence intensity of respective channel in infected and non-infected cellsTwo quality controls identify and discards outliersAll source code is available under the permissive MIT license at https://github.com/hci-unihd/batchlib. 

Impact for SARS-CoV2/COVID19 research efforts 

Develop diagnostic tools for SARS-CoV2/COVID19  

Study Type

  • In silico study / bioinformatics study 

  • In vitro study 

Strengths and limitations of the paper 

Novelty: The first study to develop a full high-throughput semi-automated imaging workflow detecting anti-SARS-CoV2 antibodies of any class with a higher specificity than ELISA 

Standing in the field:Supports the literature regarding the abundance of specific anti-SARS-Cov2 antibodies being greater at d14 post symptom onset. Uses well established image segmentation algorithms 

Appropriate statistics: Yes, the analysis algorithms are documented and well established in the field. Also, all source code is available open source under the permissive MIT license at https://github.com/hci-unihd/batchlib. 

Viral model used:VeroE6 cell infected with SARS-CoV2 

Translatability:Yes, the assay can be used for population based screening for SARS-CoV2 specific Ab presence in the serum as it offers higher specificity than ELISA 

Main limitations: comparing to ELISA: the assay requires wide-filed microscope compatible for high-throughput imaging-e.g. automated stage that can fit 96 well plate and a suitable computer for image analysis. The whole assay is 3 days long and it requires BSTL3 and maintenance of SARS-CoV2 stocks.