Reliable, robust, large-scale molecular testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential for monitoring the ongoing coronavirus disease 2019 (COVID-19) pandemic. We have developed a scalable analytical approach to detect viral proteins based on peptide immuno-affinity enrichment combined with liquid chromatography-mass spectrometry (LC-MS). This is a multiplexed strategy, based on targeted proteomics analysis and read-out by LC-MS, capable of precisely quantifying and confirming the presence of SARS-CoV-2 in phosphate-buffered saline (PBS) swab media from combined throat/nasopharynx/saliva samples. The results reveal that the levels of SARS-CoV-2 measured by LC-MS correlate well with their correspondingreal-time polymerase chain reaction (RT-PCR) read-out (r = 0.79). The analytical workflow shows similar turnaround times as regular RT-PCR instrumentation with a quantitative read-out of viral proteins corresponding to cycle thresholds (Ct) equivalents ranging from 21 to 34. Using RT-PCR as a reference, we demonstrate that the LC-MS-based method has 100% negative percent agreement (estimated specificity) and 95% positive percent agreement (estimated sensitivity) when analyzing clinical samples collected from asymptomatic individuals with a Ct within the limit of detection of the mass spectrometer (Ct ≤ 30). These results suggest that a scalable analytical method based on LC-MS has a place in future pandemic preparedness centers to complement current virus detection technologies.
【저자키워드】 COVID-19, SARS CoV-2, proteomics, mass spectrometry, Human, diagnostics, SISCAPA, 【초록키워드】 coronavirus disease, SARS-CoV-2, Coronavirus disease 2019, coronavirus, mass spectrometry, pandemic, peptide, media, complement, Viral proteins, severe acute respiratory syndrome Coronavirus, RT-PCR, polymerase chain reaction, clinical samples, sensitivity, specificity, Asymptomatic individuals, Liquid chromatography, cycle threshold, virus detection, limit of detection, Swab, molecular, respiratory, Quantitative, LC-MS, turnaround time, Analysis, Chain Reaction, phosphate, acute respiratory syndrome, phosphate-buffered saline, Viral protein, mass spectrometer, analytical method, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, positive, approach, asymptomatic individual, polymerase chain, robust, clinical sample, detect, collected, PBS, 【제목키워드】 SARS-COV-2 infection, Rapid, Quantitative, Analysis,