Rapid and sensitive detection of SARS-CoV-2 infection using quantitative peptide enrichment LC-MS analysis
© 2021, Hober et al..
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.
Errataetall: | |
---|---|
Medienart: |
E-Artikel |
Erscheinungsjahr: |
2021 |
---|---|
Erschienen: |
2021 |
Enthalten in: |
Zur Gesamtaufnahme - volume:10 |
---|---|
Enthalten in: |
eLife - 10(2021) vom: 08. Nov. |
Sprache: |
Englisch |
---|
Beteiligte Personen: |
Hober, Andreas [VerfasserIn] |
---|
Links: |
---|
Themen: |
COVID-19 |
---|
Anmerkungen: |
Date Completed 03.12.2021 Date Revised 04.04.2024 published: Electronic CommentIn: Elife. 2021 Dec 10;10:. - PMID 34889738 Citation Status MEDLINE |
---|
doi: |
10.7554/eLife.70843 |
---|
funding: |
|
---|---|
Förderinstitution / Projekttitel: |
|
PPN (Katalog-ID): |
NLM332857409 |
---|
LEADER | 01000caa a22002652 4500 | ||
---|---|---|---|
001 | NLM332857409 | ||
003 | DE-627 | ||
005 | 20240404232807.0 | ||
007 | cr uuu---uuuuu | ||
008 | 231225s2021 xx |||||o 00| ||eng c | ||
024 | 7 | |a 10.7554/eLife.70843 |2 doi | |
028 | 5 | 2 | |a pubmed24n1364.xml |
035 | |a (DE-627)NLM332857409 | ||
035 | |a (NLM)34747696 | ||
035 | |a (PII)e70843 | ||
040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
041 | |a eng | ||
100 | 1 | |a Hober, Andreas |e verfasserin |4 aut | |
245 | 1 | 0 | |a Rapid and sensitive detection of SARS-CoV-2 infection using quantitative peptide enrichment LC-MS analysis |
264 | 1 | |c 2021 | |
336 | |a Text |b txt |2 rdacontent | ||
337 | |a ƒaComputermedien |b c |2 rdamedia | ||
338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
500 | |a Date Completed 03.12.2021 | ||
500 | |a Date Revised 04.04.2024 | ||
500 | |a published: Electronic | ||
500 | |a CommentIn: Elife. 2021 Dec 10;10:. - PMID 34889738 | ||
500 | |a Citation Status MEDLINE | ||
520 | |a © 2021, Hober et al. | ||
520 | |a 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 | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a COVID-19 | |
650 | 4 | |a SARS CoV-2 | |
650 | 4 | |a SISCAPA | |
650 | 4 | |a diagnostics | |
650 | 4 | |a human | |
650 | 4 | |a immunology | |
650 | 4 | |a infectious disease | |
650 | 4 | |a inflammation | |
650 | 4 | |a mass spectrometry | |
650 | 4 | |a microbiology | |
650 | 4 | |a proteomics | |
650 | 7 | |a Peptide Fragments |2 NLM | |
650 | 7 | |a Viral Proteins |2 NLM | |
700 | 1 | |a Tran-Minh, Khue Hua |e verfasserin |4 aut | |
700 | 1 | |a Foley, Dominic |e verfasserin |4 aut | |
700 | 1 | |a McDonald, Thomas |e verfasserin |4 aut | |
700 | 1 | |a Vissers, Johannes Pc |e verfasserin |4 aut | |
700 | 1 | |a Pattison, Rebecca |e verfasserin |4 aut | |
700 | 1 | |a Ferries, Samantha |e verfasserin |4 aut | |
700 | 1 | |a Hermansson, Sigurd |e verfasserin |4 aut | |
700 | 1 | |a Betner, Ingvar |e verfasserin |4 aut | |
700 | 1 | |a Uhlén, Mathias |e verfasserin |4 aut | |
700 | 1 | |a Razavi, Morteza |e verfasserin |4 aut | |
700 | 1 | |a Yip, Richard |e verfasserin |4 aut | |
700 | 1 | |a Pope, Matthew E |e verfasserin |4 aut | |
700 | 1 | |a Pearson, Terry W |e verfasserin |4 aut | |
700 | 1 | |a Andersson, Leigh N |e verfasserin |4 aut | |
700 | 1 | |a Bartlett, Amy |e verfasserin |4 aut | |
700 | 1 | |a Calton, Lisa |e verfasserin |4 aut | |
700 | 1 | |a Alm, Jessica J |e verfasserin |4 aut | |
700 | 1 | |a Engstrand, Lars |e verfasserin |4 aut | |
700 | 1 | |a Edfors, Fredrik |e verfasserin |4 aut | |
773 | 0 | 8 | |i Enthalten in |t eLife |d 2012 |g 10(2021) vom: 08. Nov. |w (DE-627)NLM221831460 |x 2050-084X |7 nnns |
773 | 1 | 8 | |g volume:10 |g year:2021 |g day:08 |g month:11 |
856 | 4 | 0 | |u http://dx.doi.org/10.7554/eLife.70843 |3 Volltext |
912 | |a GBV_USEFLAG_A | ||
912 | |a GBV_NLM | ||
951 | |a AR | ||
952 | |d 10 |j 2021 |b 08 |c 11 |