Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Development and validation of a quantitative method for the enumeration of <i>Salmonella enterica</i> serovar Infantis from environmental poultry feces based on most probable number approach followed by confirmatory qPCR.

Frontiers in microbiology·2026

Same author

Evaluation of an E2-based indirect ELISA for the serological differentiation of ovine Italy pestivirus from classical swine fever virus in pigs.

Frontiers in veterinary science·2026

Same author

Candidate Faecal microRNAs as Non-Invasive Biomarkers for Bovine Paratuberculosis in <i>Marchigiana</i> Beef Cattle.

International journal of molecular sciences·2026

Same author

Molecular detection and characterization of Hepatitis E virus in wild boars (Sus scrofa) from northern Italy.

Research in veterinary science·2026

Same author

Feral pigeons as reservoirs of zoonotic pathogens in the urban environment: Spatial clustering and molecular evidence.

Veterinary microbiology·2026

Same author

Genetic diversity and circulation of influenza A viruses in Italian pig farms: insights from surveillance and vaccination.

Porcine health management·2026

Search research articles

Related Experiment Video

Updated: Mar 7, 2026

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

Published on: July 9, 2012

A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything.

Petr Kralik¹, Matteo Ricchi²

¹Department of Food and Feed Safety, Veterinary Research Institute Brno, Czechia.

Frontiers in Microbiology

|February 18, 2017

Summary

Quantitative PCR (qPCR) is a vital tool for detecting microbes in diagnostics and food safety. This guide clarifies qPCR principles, data interpretation, and performance parameters for users.

Keywords:

accuracy limit of detection limit of quantification precision quantitative PCR trueness

More Related Videos

On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

Published on: February 23, 2018

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Related Experiment Videos

Last Updated: Mar 7, 2026

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

Published on: July 9, 2012

On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System

Published on: February 23, 2018

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Area of Science:

Molecular Biology
Microbiology
Biotechnology

Background:

Real-time PCR (qPCR) is a standard technique for microbial detection and quantification.
Applications span clinical diagnostics, veterinary diagnostics, and food safety.
Understanding qPCR nuances is crucial for accurate results.

Purpose of the Study:

To provide a basic guide for qPCR users.
To clarify common issues in qPCR methodology and data interpretation.
To orient beginners in the application of quantitative PCR.

Main Methods:

Review of established real-time PCR principles.
Discussion of terminology, definitions, and data presentation.
Overview of qPCR performance parameters and limitations.

Main Results:

Identified key areas requiring attention for effective qPCR implementation.
Highlighted challenges in data interpretation and presentation.
Emphasized the importance of understanding qPCR limitations.

Conclusions:

A clear understanding of qPCR principles enhances diagnostic and safety applications.
Proper attention to terminology, data, and performance is essential for reliable microbial analysis.
This guide serves as a foundational resource for qPCR practitioners.