Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples
Abstract Background: SARS-CoV-2 has been found in the heart of COVID-19 patients. It is unclear how the virus passes from the upper respiratory tract to the myocardium. We hypothesized that SARS-CoV-2 is present in the blood of COVID-19 infected patients, spreading to other organs such as heart....
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2021-01-01
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| Series: | Journal of Clinical and Translational Science |
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| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866121008566/type/journal_article |
| _version_ | 1811156598825943040 |
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| author | Han-Gang Yu Gina Sizemore Katy Smoot Peter Perrotta |
| author_facet | Han-Gang Yu Gina Sizemore Katy Smoot Peter Perrotta |
| author_sort | Han-Gang Yu |
| collection | DOAJ |
| description |
Abstract
Background:
SARS-CoV-2 has been found in the heart of COVID-19 patients. It is unclear how the virus passes from the upper respiratory tract to the myocardium. We hypothesized that SARS-CoV-2 is present in the blood of COVID-19 infected patients, spreading to other organs such as heart.
Methods:
We targeted two viroporins, Orf3a and E, in SARS-CoV-2. Orf3a and E form non-voltage-gated ion channels. A combined fluorescence potassium ion assay with three channel modulators (4-aminopyridine, emodin-Orf3a channel blocker, and gliclazide-E channel blocker) was developed to detect SARS-CoV-2 Orf3a/E channel activity. In blood samples, we subtracted the fluorescence signals in the absence and presence of emodin/gliclazide to detect Orf3a and E channel activity.
Results:
In lentivirus-spiked samples, we detected significant channel activity of Orf3a/E based on increase in fluorescence induced by 4-aminopyridine, and this increase in fluorescence was inhibited by emodin and gliclazide. In 18 antigen/PCR-positive samples, our test results found 15 are positive, demonstrating 83.3% concordance. In 24 antigen/PCR-negative samples, our test results found 21 are negative, showing 87.5% concordance.
Conclusions:
We developed a cell-free test that can detect Orf3a/E channel activity of SARS-CoV-2 in blood samples from COVID-19-infected individuals, confirming a hypothesis that the virus spreads to the heart via blood circulation.
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| first_indexed | 2024-04-10T04:54:03Z |
| format | Article |
| id | doaj.art-b2dd9753260a4d209451ef5b0cbe37a6 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-10T04:54:03Z |
| publishDate | 2021-01-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-b2dd9753260a4d209451ef5b0cbe37a62023-03-09T12:31:03ZengCambridge University PressJournal of Clinical and Translational Science2059-86612021-01-01510.1017/cts.2021.856Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samplesHan-Gang Yu0https://orcid.org/0000-0001-6838-8310Gina Sizemore1https://orcid.org/0000-0002-0586-9229Katy Smoot2Peter Perrotta3Department of Physiology and Pharmacology, School of Medicine, West Virginia University, Morgantown, WV, USAEZCARE Walk-In Medical Center, Moorefield, WV, USADepartment of Pathology, Anatomy and Laboratory Medicine, School of Medicine, West Virginia University, Morgantown, WV, USADepartment of Pathology, Anatomy and Laboratory Medicine, School of Medicine, West Virginia University, Morgantown, WV, USA Abstract Background: SARS-CoV-2 has been found in the heart of COVID-19 patients. It is unclear how the virus passes from the upper respiratory tract to the myocardium. We hypothesized that SARS-CoV-2 is present in the blood of COVID-19 infected patients, spreading to other organs such as heart. Methods: We targeted two viroporins, Orf3a and E, in SARS-CoV-2. Orf3a and E form non-voltage-gated ion channels. A combined fluorescence potassium ion assay with three channel modulators (4-aminopyridine, emodin-Orf3a channel blocker, and gliclazide-E channel blocker) was developed to detect SARS-CoV-2 Orf3a/E channel activity. In blood samples, we subtracted the fluorescence signals in the absence and presence of emodin/gliclazide to detect Orf3a and E channel activity. Results: In lentivirus-spiked samples, we detected significant channel activity of Orf3a/E based on increase in fluorescence induced by 4-aminopyridine, and this increase in fluorescence was inhibited by emodin and gliclazide. In 18 antigen/PCR-positive samples, our test results found 15 are positive, demonstrating 83.3% concordance. In 24 antigen/PCR-negative samples, our test results found 21 are negative, showing 87.5% concordance. Conclusions: We developed a cell-free test that can detect Orf3a/E channel activity of SARS-CoV-2 in blood samples from COVID-19-infected individuals, confirming a hypothesis that the virus spreads to the heart via blood circulation. https://www.cambridge.org/core/product/identifier/S2059866121008566/type/journal_articleCOVID-19SARS-CoV-2channel activity detectionblood |
| spellingShingle | Han-Gang Yu Gina Sizemore Katy Smoot Peter Perrotta Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples Journal of Clinical and Translational Science COVID-19 SARS-CoV-2 channel activity detection blood |
| title | Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples |
| title_full | Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples |
| title_fullStr | Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples |
| title_full_unstemmed | Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples |
| title_short | Detecting SARS-CoV-2 Orf3a and E ion channel activity in COVID-19 blood samples |
| title_sort | detecting sars cov 2 orf3a and e ion channel activity in covid 19 blood samples |
| topic | COVID-19 SARS-CoV-2 channel activity detection blood |
| url | https://www.cambridge.org/core/product/identifier/S2059866121008566/type/journal_article |
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