Real-Time Polymerase Chain Reaction

Roche Real Time Polymerase Chain Reaction also known as quantitative Polymerase Chain Reaction, is a laboratory technique that combines reverse transcription of RNA into DNA and amplification of specific DNA targets using polymerase chain reaction. It is primarily used to measure the amount of a specific RNA. This is achieved by monitoring the amplification reaction using fluorescence, a technique called real-time PCR or quantitative PCR.

How Real Time PCR works

Roche Real time Polymerase Chain Reaction uses a nuclear-derived method for detecting the presence of specific genetic material in any pathogen, including a virus. This is how real time RT–PCR became one of the most accurate laboratory methods for detecting, tracking and studying the COVID-19 coronavirus. While many countries have used real time RT–PCR for diagnosing other diseases, such Ebola virus and Zika virus.

In order for viruses like the COVID-19 virus to be detected early in the body using real time RT–PCR, scientists need to convert the RNA to DNA. This is a process called ‘reverse transcription’. They do this because only DNA can be copied — or amplified — which is a key part of the real time.


RT–PCR process for detecting viruses

Roche Light Cylcer Real-Time Polymerase Chain Reaction checks for the presence of viral components required for the virus to reproduce and can confirmwhether a person is really infected with a virus like SARS-CoV-2 (the virus that is responsible for COVID-19).

For this to occur, the SARS-CoV-2 genetic material, which comes in the form of RNA, a single-stranded molecule, has to be converted to DNAfirst, which is a double-stranded molecule. This is done through a process called reverse transcription, which is an important part of the RT-PCR test.

With the use of RT-PCR, the result can be seen while the test is in progress.

Each sample in the RT-PCR machine is exposed to a light source and the resulting fluorescence of each reaction is measured as RT-PCR progresses.

For a sample to be recorded as positive, fluorescence measurements should be above the background fluorescence threshold.

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