How to Avoid False-Negative and False-Positive COVID-19 PCR Testing
Background: Up to 40% of test results for COVID-19 in the presence of clinical manifestations of the disease might be negative. The reason for a false-negative result might originate from any step of the analysis: poor-quality or empty swab, poor RNA isolation, inactivation of reverse transcriptase...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | International Journal of Translational Medicine |
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| Online Access: | https://www.mdpi.com/2673-8937/2/2/18 |
| _version_ | 1797486310659719168 |
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| author | Irina Fevraleva Olga Glinshchikova Tatiana Makarik Andrey Sudarikov |
| author_facet | Irina Fevraleva Olga Glinshchikova Tatiana Makarik Andrey Sudarikov |
| author_sort | Irina Fevraleva |
| collection | DOAJ |
| description | Background: Up to 40% of test results for COVID-19 in the presence of clinical manifestations of the disease might be negative. The reason for a false-negative result might originate from any step of the analysis: poor-quality or empty swab, poor RNA isolation, inactivation of reverse transcriptase or Taq polymerase in the test. Methods: Here we describe a PCR approach for SARS-CoV-2 detection with swab quality and integrity controlled by human <i>ABL1</i> mRNA amplification. Designed primers work with the cDNA of the <i>ABL1</i> gene, not genomic DNA. Results: The simultaneous appearance of three signals corresponding to the nucleocapsid, spike, and <i>ABL1</i> gene indicates infection with the Omicron strain. The amplification of <i>ABL1</i> gene and nucleocapsid only indicate other than Omicron infection. The appearance of ABL1 amplification only indicates a true negative result for SARS-CoV-2. All other variants are null and void. Conclusions: A system has been developed for multiplex PCR diagnostics of SARS-CoV-2, which makes it possible to eliminate errors leading to false-negative and false-positive results at all stages of analysis. This is accomplished by the presence of specific primers for human RNA, controlling proper swab application, handling, and all the stages of RT-PCR. |
| first_indexed | 2024-03-09T23:31:22Z |
| format | Article |
| id | doaj.art-a07cd55bc3e541268415b9ab5338489c |
| institution | Directory Open Access Journal |
| issn | 2673-8937 |
| language | English |
| last_indexed | 2024-03-09T23:31:22Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Translational Medicine |
| spelling | doaj.art-a07cd55bc3e541268415b9ab5338489c2023-11-23T17:08:47ZengMDPI AGInternational Journal of Translational Medicine2673-89372022-06-012220420910.3390/ijtm2020018How to Avoid False-Negative and False-Positive COVID-19 PCR TestingIrina Fevraleva0Olga Glinshchikova1Tatiana Makarik2Andrey Sudarikov3National Medical Research Center for Hematology, 125167 Moscow, RussiaNational Medical Research Center for Hematology, 125167 Moscow, RussiaNational Medical Research Center for Hematology, 125167 Moscow, RussiaNational Medical Research Center for Hematology, 125167 Moscow, RussiaBackground: Up to 40% of test results for COVID-19 in the presence of clinical manifestations of the disease might be negative. The reason for a false-negative result might originate from any step of the analysis: poor-quality or empty swab, poor RNA isolation, inactivation of reverse transcriptase or Taq polymerase in the test. Methods: Here we describe a PCR approach for SARS-CoV-2 detection with swab quality and integrity controlled by human <i>ABL1</i> mRNA amplification. Designed primers work with the cDNA of the <i>ABL1</i> gene, not genomic DNA. Results: The simultaneous appearance of three signals corresponding to the nucleocapsid, spike, and <i>ABL1</i> gene indicates infection with the Omicron strain. The amplification of <i>ABL1</i> gene and nucleocapsid only indicate other than Omicron infection. The appearance of ABL1 amplification only indicates a true negative result for SARS-CoV-2. All other variants are null and void. Conclusions: A system has been developed for multiplex PCR diagnostics of SARS-CoV-2, which makes it possible to eliminate errors leading to false-negative and false-positive results at all stages of analysis. This is accomplished by the presence of specific primers for human RNA, controlling proper swab application, handling, and all the stages of RT-PCR.https://www.mdpi.com/2673-8937/2/2/18COVID-19SARS-CoV-2RT-PCR |
| spellingShingle | Irina Fevraleva Olga Glinshchikova Tatiana Makarik Andrey Sudarikov How to Avoid False-Negative and False-Positive COVID-19 PCR Testing International Journal of Translational Medicine COVID-19 SARS-CoV-2 RT-PCR |
| title | How to Avoid False-Negative and False-Positive COVID-19 PCR Testing |
| title_full | How to Avoid False-Negative and False-Positive COVID-19 PCR Testing |
| title_fullStr | How to Avoid False-Negative and False-Positive COVID-19 PCR Testing |
| title_full_unstemmed | How to Avoid False-Negative and False-Positive COVID-19 PCR Testing |
| title_short | How to Avoid False-Negative and False-Positive COVID-19 PCR Testing |
| title_sort | how to avoid false negative and false positive covid 19 pcr testing |
| topic | COVID-19 SARS-CoV-2 RT-PCR |
| url | https://www.mdpi.com/2673-8937/2/2/18 |
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