An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay

Jing Zou, Chaitanya Kurhade, Hope C. Chang, Yanping Hu, Jose A. Meza, David Beaver, Ky Trinh, Joseph Omlid, Bassem Elghetany, Ragini Desai, Peter McCaffrey, Juan D. Garcia, Pei-Yong Shi, Ping Ren, Xuping Xie

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


A reliable and efficient serological test is crucial for monitoring neutralizing antibodies against SARS-CoV-2 and its variants of concern (VOCs). Here, we present an integrated research–clinical platform for a live SARS-CoV-2 neutralization assay, utilizing highly attenuated SARS-CoV-2 (Δ3678_WA1-spike). This strain contains mutations in viral transcription regulation sequences and deletion in the open-reading-frames 3, 6, 7, and 8, allowing for safe handling in biosafety level 2 (BSL-2) laboratories. Building on this backbone, we constructed a genetically stable reporter virus (mGFP Δ3678_WA1-spike) by incorporating a modified green fluorescent protein sequence (mGFP). We also constructed mGFP Δ3678_BA.5-spike and mGFP Δ3678_XBB.1.5-spike by substituting the WA1 spike with variants BA.5 and XBB.1.5 spike, respectively. All three viruses exhibit robust fluorescent signals in infected cells and neutralization titers in an optimized fluorescence reduction neutralization assay that highly correlates with a conventional plaque reduction assay. Furthermore, we established that a streamlined robot-aided Bench-to-Clinics COVID-19 Neutralization Test workflow demonstrated remarkably sensitive, specific, reproducible, and accurate characteristics, allowing the assessment of neutralization titers against SARS-CoV-2 variants within 24 h after sample receiving. Overall, our innovative approach provides a valuable avenue for large-scale testing of clinical samples against SARS-CoV-2 and VOCs at BSL-2, supporting pandemic preparedness and response strategies.

Original languageEnglish (US)
Article number1855
Issue number9
StatePublished - Sep 2023


  • COVID-19
  • SARS-CoV-2
  • high throughput
  • live attenuated
  • neutralization assay
  • serological diagnosis
  • variants of concern

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology


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