Definition and research guide
This page combines a MeSH definition, publication statistics and selected research abstracts. It is a starting point for reading the literature, not a medical textbook or a systematic evidence review.
Content available: 4 complete abstracts (850 words), 10 references. Full article text is not stored or reproduced here; follow the PMID, DOI or PMC links to read the original sources and their access terms.
Definition
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
Definition: NLM Medical Subject Headings (MeSH) scope note (Public domain (U.S. National Library of Medicine); attribution requested). View the MeSH record.
Also known as: Polymerase Chain Reaction, Reverse Transcriptase; Reverse Transcriptase PCR; PCR, Reverse Transcriptase; Transcriptase PCR, Reverse.
Where it fits in the MeSH classification
- Broader topics
- Polymerase Chain Reaction
- See also
- RNA-Directed DNA Polymerase
Research landscape
PubMed indexes 155,092 records tagged with the MeSH heading “Reverse Transcriptase Polymerase Chain Reaction” (counts retrieved 2026-10-29). 3,700 of them (2.4%) were published from 2020 onward, and the busiest period so far is 2000-2009 with 91,195 records.
Reviews account for 1,592 records (1.0%) and meta-analyses for 119. Randomized controlled trials number 547 (0.4%) as a share of all records on the topic. Randomized trials make up only a small share of this literature, so much of what is known comes from laboratory, observational or descriptive work.
Show the data
| Period | Records |
|---|---|
| through 1999 | 4,683 |
| 2000-2009 | 91,195 |
| 2010-2019 | 55,514 |
| 2020 onward | 3,700 |
Publication-type chart data
| Type | Records | Share |
|---|---|---|
| Reviews | 1,592 | 1.0% |
| Meta-analyses | 119 | 0.1% |
| Randomized controlled trials | 547 | 0.4% |
Method: PubMed query "Reverse Transcriptase Polymerase Chain Reaction"[MeSH Terms] with publication-type and date filters, retrieved 2026-10-29.
Reading the evidence in genetics and genomics
Genetic findings vary widely in strength. A variant linked to a condition in one study may not replicate in other ancestries, and most common variants have small effects that are meaningful only in aggregate. Studies of rare, high-impact variants can support strong causal claims about a gene, while association studies describe statistical links that need functional and independent replication. Check the sample size, the ancestry of participants, whether results were replicated, and whether a laboratory or clinical outcome was actually measured.
Evidence from open-access studies
These open-access studies come from PubMed’s best-match results for this heading (papers whose main topic it is), ranked here by citation count, with one recent addition. Each abstract is reproduced in full under its recorded open licence so that you can read the authors’ own words. They illustrate the literature; they are not a systematic summary of it. Citation counts measure attention, not reliability or clinical benefit.
| Study / authors | Year | Source label / design | Europe PMC citations | Licence / source |
|---|---|---|---|---|
| Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR | 2020 | Research article Design not recorded in structured metadata | 359 | cc by PMID 32968075 · DOI · PMC7511968 |
| Rapid isothermal amplification and portable detection system for SARS-CoV-2 | 2020 | Research article Design not recorded in structured metadata | 315 | cc by PMID 32868442 · DOI · PMC7502724 |
| Estimating the false-negative test probability of SARS-CoV-2 by RT-PCR | 2020 | Review article Design not recorded in structured metadata | 197 | cc by PMID 33334398 · DOI · PMC7812420 |
| Diagnostic value of RT-PCR for detection of tick-borne encephalitis virus RNA in cerebrospinal fluid | 2026 | Research article Design not recorded in structured metadata | 0 | cc by PMID 41526762 · DOI · PMC13086654 |
Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR [1]
Abstract reading summary — source excerpts
- Opening excerpt
- Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing.
- Closing excerpt
- This could aid expansion of COVID-19 testing.
- Interpretation limits
- Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (150 words)
Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing. Methodological simplification could increase diagnostic availability and efficiency, benefitting patient care and infection control. Here, we describe methods circumventing RNA extraction in COVID-19 testing by performing RT-PCR directly on heat-inactivated or lysed samples. Our data, including benchmarking using 597 clinical patient samples and a standardised diagnostic system, demonstrate that direct RT-PCR is viable option to extraction-based tests. Using controlled amounts of active SARS-CoV-2, we confirm effectiveness of heat inactivation by plaque assay and evaluate various generic buffers as transport medium for direct RT-PCR. Significant savings in time and cost are achieved through RNA-extraction-free protocols that are directly compatible with established PCR-based testing pipelines. This could aid expansion of COVID-19 testing.
Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 32968075 · DOI · PMC7511968
Rapid isothermal amplification and portable detection system for SARS-CoV-2 [2]
Abstract reading summary — source excerpts
- Opening excerpt
- The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs.
- Closing excerpt
- This work demonstrates an alternative pathway for SARS-CoV-2 diagnostics that does not require conventional laboratory infrastructure, in settings where diagnosis is required at the point of sample collection.
- Interpretation limits
- Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (246 words)
The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs. As assay protocols and primer sequences become widely known, many laboratories perform diagnostic tests using methods such as RT-PCR or reverse transcription loop mediated isothermal amplification (RT-LAMP). Here, we report an RT-LAMP isothermal assay for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus and demonstrate the assay on clinical samples using a simple and accessible point-of-care (POC) instrument. We characterized the assay by dipping swabs into synthetic nasal fluid spiked with the virus, moving the swab to viral transport medium (VTM), and sampling a volume of the VTM to perform the RT-LAMP assay without an RNA extraction kit. The assay has a limit of detection (LOD) of 50 RNA copies per μL in the VTM solution within 30 min. We further demonstrate our assay by detecting SARS-CoV-2 viruses from 20 clinical samples. Finally, we demonstrate a portable and real-time POC device to detect SARS-CoV-2 from VTM samples using an additively manufactured three-dimensional cartridge and a smartphone-based reader. The POC system was tested using 10 clinical samples, and was able to detect SARS-CoV-2 from these clinical samples by distinguishing positive samples from negative samples after 30 min. The POC tests are in complete agreement with RT-PCR controls. This work demonstrates an alternative pathway for SARS-CoV-2 diagnostics that does not require conventional laboratory infrastructure, in settings where diagnosis is required at the point of sample collection.
Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 32868442 · DOI · PMC7502724
Estimating the false-negative test probability of SARS-CoV-2 by RT-PCR [3]
Abstract reading summary — source excerpts
- Opening excerpt
- BackgroundReverse-transcription PCR (RT-PCR) assays are used to test for infection with the SARS-CoV-2 virus.
- Closing excerpt
- This has implications for identifying infected patients, contact tracing and discharging convalescing patients who are potentially still infectious.
- Interpretation limits
- Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (223 words)
BackgroundReverse-transcription PCR (RT-PCR) assays are used to test for infection with the SARS-CoV-2 virus. RT-PCR tests are highly specific and the probability of false positives is low, but false negatives are possible depending on swab type and time since symptom onset.AimTo determine how the probability of obtaining a false-negative test in infected patients is affected by time since symptom onset and swab type.MethodsWe used generalised additive mixed models to analyse publicly available data from patients who received multiple RT-PCR tests and were identified as SARS-CoV-2 positive at least once.ResultsThe probability of a positive test decreased with time since symptom onset, with oropharyngeal (OP) samples less likely to yield a positive result than nasopharyngeal (NP) samples. The probability of incorrectly identifying an uninfected individual due to a false-negative test was considerably reduced if negative tests were repeated 24 hours later. For a small false-positive test probability (<0.5%), the true number of infected individuals was larger than the number of positive tests. For a higher false-positive test probability, the true number of infected individuals was smaller than the number of positive tests.ConclusionNP samples are more sensitive than OP samples. The later an infected individual is tested after symptom onset, the less likely they are to test positive. This has implications for identifying infected patients, contact tracing and discharging convalescing patients who are potentially still infectious.
Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 33334398 · DOI · PMC7812420
Diagnostic value of RT-PCR for detection of tick-borne encephalitis virus RNA in cerebrospinal fluid [4]
Abstract reading summary — source excerpts
- Opening excerpt
- PURPOSE: Tick-borne encephalitis (TBE) diagnosis is mainly based on serology, which however can be difficult to interpret in previously TBE-vaccinate patients and in immunosuppressed patients.
- Closing excerpt
- The presence of TBEV RNA in CSF may correlate to severe infection and an increased risk of fatal outcome.
- Interpretation limits
- Verbatim excerpts from the authors’ abstract; the closing excerpt may describe a finding, limitation or recommendation. Design, population, effect size and uncertainty have not been independently extracted or appraised. Read the complete abstract and linked paper before interpreting the findings.
Read complete abstract (231 words)
PURPOSE: Tick-borne encephalitis (TBE) diagnosis is mainly based on serology, which however can be difficult to interpret in previously TBE-vaccinate patients and in immunosuppressed patients. The objectives of this study were to investigate if the detection of tick-borne encephalitis virus (TBEV) RNA in cerebrospinal fluid (CSF) can be used to diagnose TBE, and to assess the relation of TBEV RNA positivity to clinical characteristics and disease severity. METHODS: A retrospective study was conducted in TBE patients from Region Stockholm and Uppsala, Sweden, 2015–2020. CSF-samples were analysed for TBEV RNA using RT-PCR. Data on clinical parameters, retrieved from medical records, were analysed based on TBEV RNA positivity in CSF. RESULTS: CSF from 36 TBE patients were analysed for the presence of TBEV RNA. TBEV RNA was detected in twenty of these patients (56%). Patients with detectable TBEV RNA in CSF had a more severe disease, with a higher rate of intensive care and assisted ventilation treatment. Overall, the mortality was 40% in patients with detectable TBEV RNA in CSF. In a subgroup of 8 patients with immunosuppressive treatment and detectable TBEV RNA in CSF, 7 (88%) had a fatal outcome. CONCLUSION: TBEV RT-PCR can be used as a complement for diagnostics in patients with underlying comorbidities involving the immune system or immunosuppressive treatment. The presence of TBEV RNA in CSF may correlate to severe infection and an increased risk of fatal outcome.
Abstract reproduced from the open-access article under the recorded cc by licence; © the authors. PMID 41526762 · DOI · PMC13086654
Key reviews and meta-analyses
The most cited review-type papers among PubMed’s best-match results for this heading, listed with bibliographic details only. Follow the links for the full text and access terms.
- Research article Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [5]
- Research article Real-time RT-PCR normalisation; strategies and considerations [6]
- Research article Standardization and quality control studies of 'real-time' quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program [7]
- Research article Real-time PCR for mRNA quantitation [8]
- Research article Variation in False-Negative Rate of Reverse Transcriptase Polymerase Chain Reaction-Based SARS-CoV-2 Tests by Time Since Exposure [9]
- Research article An overview of real-time quantitative PCR: applications to quantify cytokine gene expression [10]
References
- Smyrlaki I, Ekman M, Lentini A, Rufino de Sousa N, Papanicolaou N, Vondracek M, et al.. Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR. . 2020. PMID 32968075 · DOI · PMC7511968
- Ganguli A, Mostafa A, Berger J, Aydin MY, Sun F, Ramirez SAS, et al.. Rapid isothermal amplification and portable detection system for SARS-CoV-2. . 2020. PMID 32868442 · DOI · PMC7502724
- Wikramaratna PS, Paton RS, Ghafari M, Lourenço J. Estimating the false-negative test probability of SARS-CoV-2 by RT-PCR. . 2020. PMID 33334398 · DOI · PMC7812420
- Glans H, Bartholdsson S, Lagerqvist N, Nord H, Blennow O, Morén J, et al.. Diagnostic value of RT-PCR for detection of tick-borne encephalitis virus RNA in cerebrospinal fluid. . 2026. PMID 41526762 · DOI · PMC13086654
- Bustin SA. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000. PMID 11013345 · DOI
- Huggett J, Dheda K, Bustin S, Zumla A. Real-time RT-PCR normalisation; strategies and considerations. Genes Immun. 2005. PMID 15815687 · DOI
- Gabert J, Beillard E, van der Velden VH, Bi W, Grimwade D, Pallisgaard N, et al.. Standardization and quality control studies of 'real-time' quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia - a Europe Against Cancer program. Leukemia. 2003. PMID 14562125 · DOI
- Wong ML, Medrano JF. Real-time PCR for mRNA quantitation. Biotechniques. 2005. PMID 16060372 · DOI
- Kucirka LM, Lauer SA, Laeyendecker O, Boon D, Lessler J. Variation in False-Negative Rate of Reverse Transcriptase Polymerase Chain Reaction-Based SARS-CoV-2 Tests by Time Since Exposure. Ann Intern Med. 2020. PMID 32422057 · DOI · PMC7240870
- Giulietti A, Overbergh L, Valckx D, Decallonne B, Bouillon R, Mathieu C. An overview of real-time quantitative PCR: applications to quantify cytokine gene expression. Methods. 2001. PMID 11846608 · DOI
About this article
- Sources
- NLM Medical Subject Headings (definition, synonyms); NCBI PubMed (counts, identifiers); Europe PMC (open-access abstracts and citation counts).
- Verification
- Every reference was resolved against PubMed on 2026-10-31; title, year and DOI matched.
- Review state
- automated source verified; no human expert review has been recorded yet.
- Version
- 1, last updated 2026-10-29.
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Evidence strength
EvidenceHelix's computed view of the study designs behind this topic: how common meta-analyses, randomized trials and reviews are, how recent the literature is, and a plain-language reading. It describes the design mix of the literature. It is not a quality appraisal of any study, and it is not medical advice.