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if(isHidden(s)){ s.style.display = 'inline'; } else { s.style.display = 'none'; } } function isHidden(element){ var style = window.getComputedStyle(element); return (style.display === 'none'); } </script> <span class="cc_sidebar"> <span class="cc_color_bar"> <span class="cc_home_icon"> <a href="https://www.thermofisher.com/blog/clinical-conversations/"><img src="https://www.thermofisher.com/blog/clinical-conversations/wp-content/themes/clinical-conversations/images/icon_home.png" alt="Clinical Conversation blog - home icon" /></a> </span> <span class="cc_post_category"> <a class='cc_colorbar_category' href='https://www.thermofisher.com/blog/clinical-conversations/category/science-and-education/'>Science and education</a> </span>  <span id="cc_post_readtime"> 5 mins. </span> <span class="cc_toggle_social"> <a href="javascript:cc_toggle('cc_jetpack_shares');">Sharing ▼</a> </span> <span class="cc_toggle"> <a href="javascript:cc_toggle('cc_metadata');">Details ▼</a> </span> </span> </span> <span id="cc_jetpack_shares"> <ul> <li><a target="_blank" href='https://www.linkedin.com/shareArticle?mini=true&url=https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Fclinical-conversations%2Fwhy-qpcr-is-the-gold-standard-for-covid-19-testing%2F&title=Why+qPCR+is+the+gold+standard+for+COVID-19+testing'><img src="https://www.thermofisher.com/blog/clinical-conversations//wp-content/themes/clinical-conversations/images/social_icons_linkedin.jpg" alt="Linked In sharing icon" /></a></li> <li><a target="_blank" href='https://www.facebook.com/sharer/sharer.php?u=https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Fclinical-conversations%2Fwhy-qpcr-is-the-gold-standard-for-covid-19-testing%2F'><img src="https://www.thermofisher.com/blog/clinical-conversations//wp-content/themes/clinical-conversations/images/social_icons_facebook.jpg" alt="Facebook sharing icon" /></a></li> <li><a target="_blank" href='https://twitter.com/intent/tweet?text=Why+qPCR+is+the+gold+standard+for+COVID-19+testing&url=https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Fclinical-conversations%2Fwhy-qpcr-is-the-gold-standard-for-covid-19-testing%2F&'><img src="https://www.thermofisher.com/blog/clinical-conversations//wp-content/themes/clinical-conversations/images/social_icons_twitter.jpg" alt="Twitter sharing icon" /></a></li> <li><a href='mailto:?subject=You might be interested in Why qPCR is the gold standard for COVID-19 testing from the Clinical Conversations blog.&body=You might be interested in Why qPCR is the gold standard for COVID-19 testing from the Clinical Conversations blog at https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Fclinical-conversations%2Fwhy-qpcr-is-the-gold-standard-for-covid-19-testing%2F'><img src="https://www.thermofisher.com/blog/clinical-conversations/wp-content/themes/clinical-conversations/images/icon_email.png" alt="Email sharing icon" /></a></li> </ul> </span> <span id="cc_metadata"> <span class="cc_dateline"> <span class="post-info-date">  23 Oct 2020 </span> || <span class="post-info-author">   By Manoj Gandhi, M.D., Ph.D. </span> </span> <span class="cc_shares"> <span class="cc_shares_subhead" >Shares: <span class="share_count">7</span> </span> </span> <span class="cc_version"> <span class="cc_version_subhead">Versions of this article</span> Original article. </span> <span class="cc_post_tags"> <span class="cc_post_tags_subhead">Tags</span> <a href='https://www.thermofisher.com/blog/clinical-conversations/tag/covid-19/'>COVID-19</a>, <a href='https://www.thermofisher.com/blog/clinical-conversations/tag/qpcr/'>QPCR</a>, <a href='https://www.thermofisher.com/blog/clinical-conversations/tag/sars-cov-2/'>Sars-cov-2</a> </span>   </span> <script type="text/javascript"> cc_toggle('cc_metadata'); cc_toggle('cc_jetpack_shares'); </script> </header><div class="entry-content"><p><em>Every day there’s a new headline about the best ways to test for SARS-CoV-2 virus, the virus that causes COVID-19. In the following Q&A, Joshua Trotta, Senior Director Business Development, discusses what you need to know about qPCR, the gold standard for COVID-19 testing. This is a follow-up to our “<a href="https://www.thermofisher.com/blog/ask-a-scientist/how-does-a-covid-19-test-work/">How does a COVID-19 test work?</a>” interview with Joshua on Science with a Twist earlier this year.</em></p> <p><strong>What is an RT-qPCR test?</strong></p> <p>“PCR” is shorthand for “polymerase chain reaction,” a scientific methodology to make millions and billions of copies of a specific DNA sample, which for the past 25 years has been one of the most widely used techniques in molecular biology.</p> <p>The genetic material in SARS-CoV-2 virus is stored in RNA. In the case of COVID-19 testing, RNA from a SARS-CoV-2 sample is first converted to its complementary DNA sequence by a process called reverse transcription (RT). The DNA thus transcribed is amplified by qPCR, where “q” stands for “quantitative”; hence the technique is called RT-qPCR[1].</p> <p><img decoding="async" class="cc_image_thumbnail alignnone" style="margin: 10px 10px 10px 0" src="https://www.thermofisher.com/blog/clinical-conversations/wp-content/uploads/sites/18/2020/10/joshua-trotta.jpg" alt="Joshua Trotta" width="768" height="1024" align="left" /> RT-qPCR can detect the genetic information (RNA) of the novel coronavirus—even if the virus is present in extremely small amounts. Researchers estimate that RT-qPCR can detect as little as ~1,000 copies of viral RNA per milliliter, or 10 copies per analytical limit of detection (LoD). This means it is possible to diagnose someone as positive for COVID-19 even when their sample contains a very small amount of virus.</p> <p>This means that COVID-19 RT-qPCR tests are not only able to detect the virus’s genetic information, they are also able to quantify the amount of that genetic information that is present in a sample. The <a href="https://www.thermofisher.com/us/en/home/clinical/clinical-genomics/pathogen-detection-solutions/sars-cov-2-covid-19.html">Applied Biosystems TaqPath COVID-19 Combo Kit</a> from Thermo Fisher Scientific is designed to detect the presence of the virus at very low levels and uses an algorithm to provide a positive or negative result.</p> <p><strong>How is a SARS-CoV-2 sample collected for RT-qPCR testing? </strong></p> <p>Samples can be collected in a few different ways. To test if someone with COVID-19 symptoms is infected with SARS-CoV-2, a health official might collect cells from the person’s nose and/or throat usually by using a nasal, throat or nasopharyngeal swab. Alternatively, a saliva sample can be collected.</p> <p>As organizations look for <a href="https://www.thermofisher.com/us/en/home/clinical/clinical-genomics/pathogen-detection-solutions/covid-19-workplace-testing.html">routine testing solutions in workplaces and universities</a>, they are also exploring options for self-collection. These self-collected samples are then sent to a lab where the RT-qPCR test is run, which takes about two to four hours to complete.</p> <p><strong>What is an antigen COVID-19 test?</strong></p> <p>While an RT-qPCR COVID-19 test detects the genetic material of the novel coronavirus, an <a href="https://www.thermofisher.com/us/en/home/clinical/clinical-genomics/pathogen-detection-solutions/covid-19-sars-cov-2/pcr-antigen-testing-technologies.html">antigen-based COVID-19 test</a> is designed to detect something different: proteins on or within the virus.</p> <p>For someone getting an antigen COVID-19 test, the sample is collected in the same way as for the RT-qPCR COVID-19 test—usually by nasopharyngeal swab. However, in the case of an antigen COVID-19 test, the sample does not need to be sent to a lab for analysis. Instead, it can be analyzed at the point of care in a process that can take as little as 15 minutes.</p> <p>The trade-off for the speed and convenience of an antigen COVID-19 test is specificity. The antigen COVID-19 test requires a higher viral load in the sample to yield a positive result. As a result, someone who tests negative with an antigen test may actually have an active COVID-19 infection, but there are too few virus proteins present in the sample to register positive on the test. That same person my take an antigen test the next day and get a positive result, simply because they have more of the virus in their system.</p> <p><strong>What are the main differences between the different types of COVID-19</strong> <strong>tests?</strong></p> <p>When it comes to <a href="https://www.thermofisher.com/us/en/home/clinical/clinical-genomics/pathogen-detection-solutions/covid-19-sars-cov-2.html">COVID-19 testing</a>, your choice of test type usually is a choice between accuracy and speed. You need to fully understand the tools available in the toolkit, and then pull out the right one at the right time.</p> <p>A COVID-19 antigen test can provide results quickly, but RT-qPCR is considered the gold standard for COVID-19 testing as it provides a more definitive answer as to whether an individual has the virus in their body.</p> <p>In fact, in situations where antigen testing is used for rapid screening, negative results are often sent to the lab for confirmatory RT-qPCR testing to verify if the individual is indeed virus-free.</p> <p>In addition, there is a third type of test: a COVID-19 antibody test. An antibody test is a blood test that does not detect active virus. Instead, it detects antibodies to SARS-CoV-2. The presence of antibodies indicates that someone is actively dealing with, or recovered from, a COVID-19 infection.</p> <p><strong>What factors can lead to a false-positive or false-negative test result?</strong></p> <p>It’s also important to understand that, like any other diagnostic test, the results of COVID-19 tests can be impacted by sample collection. The reliability and accuracy of these tests depend on:</p> <ul> <li>How the sample was collected—it is always recommended to follow the manufacturer’s Instructions for Use (IFU) on the proper use of the test and approved sample collection methods.</li> <li>How many samples were collected—for nasopharyngeal and nasal swabs, samples from both nostrils are recommended.</li> <li>When the sample was collected—for example, someone who is newly infected may not have detectable amounts of virus in their system.</li> <li>How long ago the sample was taken—samples should arrive at the lab within 72 hours after collection.</li> <li>How the sample was stored before analysis—again, it is important to follow the manufacturer’s IFU, as extreme temperatures or other harsh conditions can damage viral molecular integrity.</li> </ul> <p><strong>What lies ahead for COVID-19 testing?</strong></p> <p>Health officials predict an increase in SARS-CoV-2 infections and they continue to weigh the pros and cons of different testing methodologies and protocols.</p> <p>qPCR testing is going to become even more important given its ability to detect SARS-CoV-2 even when it’s present in extremely small quantities, like when someone has been recently infected and has a relatively small viral load.</p> <p>With qPCR, it’s possible to make a diagnosis early on in the infection, meaning the infected individual can go into isolation earlier to prevent passing the virus on to others. Until a vaccine is widely available, testing is the key to stopping this crisis.</p> <p><strong>Additional resources</strong></p> <ul> <li><a href="https://www.thermofisher.com/us/en/home/clinical/clinical-genomics/pathogen-detection-solutions/covid-19-sars-cov-2.html">COVID-19 testing products</a></li> <li><a href="https://www.thermofisher.com/us/en/home/global/forms/life-science/digital-experience.html">Real-time PCR Handbook</a></li> </ul> <hr /> <p>TaqPath COVID-19 Combo Kit: For Emergency Use Authorization (EUA) only. For prescription use only. For in vitro diagnostic use.</p> <p>[1] Bustin SA, Nolan T. RT-qPCR Testing of SARS-CoV-2: A Primer. <em>Int J Mol Sci</em>. 2020;21(8):3004. 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