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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Related Experiment Video

Updated: Feb 23, 2026

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
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Comparison between RT-PCR and TMA for SARS-CoV-2 detection and proposal of a diagnostic algorithm based on cut-off

Albert Bernet Sánchez1, Alba Bellés Bellés1, Eric López González1

  • 1Microbiology Section, Hospital Universitari Arnau de Vilanova [Arnau de Vilanova University Hospital], Lleida, Spain; Institut de Recerca Biomèdica [Biomedical Research Institute] - Fundació Dr. Pifarré, IRBLleida, Lleida, Spain.

Enfermedades Infecciosas Y Microbiologia Clinica (English Ed.)
|February 21, 2026
PubMed
Summary

Transcription-mediated amplification (TMA) shows promise as a SARS-CoV-2 screening tool. Lowering the positivity threshold to 350 relative light units (RLU) and confirming with RT-PCR improves accuracy for this diagnostic method.

Keywords:
Algoritmo diagnósticoAmplificación mediada por transcripción (TMA)Cut-off valuesDiagnostic algorithmDiagnóstico molecularMolecular diagnosticsRT-PCRSARS-CoV-2Transcription-Mediated-Amplification (TMA)Valores de corte

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Area of Science:

  • Clinical diagnostics
  • Molecular biology
  • Virology

Background:

  • The COVID-19 pandemic necessitated rapid development and implementation of diagnostic tests.
  • Transcription-mediated amplification (TMA) emerged as a key technique to meet high diagnostic demand.
  • Evaluating TMA against RT-PCR in clinical settings is crucial for optimizing SARS-CoV-2 detection.

Purpose of the Study:

  • To compare the performance of TMA with RT-PCR for SARS-CoV-2 detection in routine clinical samples.
  • To analyze the correlation between relative light units (RLU) from TMA and RT-PCR results.
  • To establish potential cut-off ranges for TMA to enhance its diagnostic utility.

Main Methods:

  • A prospective study involving 15,156 clinical samples analyzed over one year.
  • Parallel testing of samples using TMA (Aptima Hologic) and RT-PCR (various platforms).
  • Calculation of concordance (κ statistic), sensitivity, and specificity, with RT-PCR as the reference standard.

Main Results:

  • Substantial concordance (κ = 0.692) between TMA and RT-PCR was observed.
  • TMA demonstrated a sensitivity of 97.89% and a specificity of 64.74%.
  • Analysis of RLU values revealed discrepancies, with TMA negatives (350-559 RLU) being RT-PCR positive in 20.56% of cases.

Conclusions:

  • TMA can be effectively utilized as a screening method for SARS-CoV-2.
  • Adjusting the positivity threshold to 350 RLU is recommended for TMA screening.
  • Confirmation of positive TMA results by RT-PCR is essential for accurate diagnosis.