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Real Time RT-PCR

Real Time RT-PCR

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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Updated: Sep 4, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification

Published on: March 31, 2021

Quantitative Real-Time RT-PCR Systems to Detect SARS-CoV-2.

Sumino Yanase^1,2, Hiroyoshi Sasahara³, Momoko Nabetani³

¹Department of Health Science, Daito Bunka University School of Sports & Health Science, Higashi-matsuyama, Saitama, Japan. syanase@ic.daito.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)

|July 15, 2022

Summary

Real-time RT-PCR testing can now detect SARS-CoV-2 RNA in blood samples, offering a new option beyond swabs. This advancement aids in controlling COVID-19 spread by identifying positive, asymptomatic, and negative cases.

Keywords:

Asymptomatic COVID-19 infection Blood sample Real-time RT-PCR SARS-CoV-2 RNA

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Last Updated: Sep 4, 2025

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Published on: March 31, 2021

Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

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Published on: February 11, 2022

Area of Science:

Medical Diagnostics
Virology
Molecular Biology

Background:

The COVID-19 pandemic necessitated widespread diagnostic testing for SARS-CoV-2.
Real-time RT-PCR using nasopharyngeal swabs became the standard diagnostic method in Japan.
Limitations existed in sample types for SARS-CoV-2 detection.

Purpose of the Study:

To introduce a clinical laboratory test for detecting SARS-CoV-2 RNA in blood samples.
To evaluate the use of serum, plasma, and whole blood for viral RNA detection.
To expand diagnostic capabilities beyond traditional swab-based methods.

Main Methods:

Utilized the High Pure viral nucleic acid kit for sample preparation.
Employed a real-time RT-PCR system for viral RNA detection.
Tested serum, plasma, and whole blood samples.

Main Results:

Successfully detected SARS-CoV-2 RNA in blood samples (serum, plasma, whole blood).
Demonstrated the feasibility of using blood as an alternative sample type for COVID-19 diagnosis.
Validated the High Pure viral nucleic acid kit and RT-PCR system for this application.

Conclusions:

Blood samples can be effectively used for SARS-CoV-2 RNA detection via real-time RT-PCR.
This method provides an alternative to nasopharyngeal swabs, potentially improving accessibility and patient comfort.
The study contributes to expanding diagnostic options for managing the COVID-19 pandemic.