What is RT PCR? |Reverse Transcriptase PCR
RT PCR or Reverse Transcriptase PCR is a test used to detect retroviral diseases like AIDS. It gained popularity during coronavirus outbreak.
Before knowing about RT PCR, please refer PCR or Polymerase chain reaction first here – https://sciencebirdie.com/2020/06/17/what-is-pcr-polymerase-chain-reaction-explained/
PCR or polymerase chain reaction is the technique in which the DNA is amplified and the billion DNA copies are made. But, it has exceptions like it can only detect only DNA viruses. Therefore, detection of RNA viruses like retroviruses cannot be possible.
This is where the RT PCR has the significance.
Viruses like Coronavirus have RNA as the genetic material, therefore, the detection using the traditional PCR becomes impossible.
So, scientists convert RNA to DNA using an enzyme called Reverse transcriptase, the process known as Reverse Transcription. On the other hand, the conversion of DNA to RNA is called Transcription.
Steps in RT PCR
Firstly, the RNA converted to DNA using reverse transcription.
The DNA then transferred to the thermocycler machine or the PCR machine.
Afterwards, the steps denaturation, annealing and extension followed and the billion DNA copies extracted.
Please refer the PCR steps here – https://sciencebirdie.com/2020/06/17/what-is-pcr-polymerase-chain-reaction-explained/
Coming to advantages, it had got more in addition than the traditional PCR.
Highly sensitive as the RNA involvement in it.
Highly specific, with more accuracy than PCR.
Requires less than 4 hours in diagnosis, moreover, the laboratories take up to 6-8 hours.
As the genetic material placed in a closed tube with enzymes, it has low potential contamination.
RT PCR in Corona test
Nowadays, with Coronavirus being prevalent, the RT PCR use got very much boosted up.
Firstly, the victim’s blood sample is taken then using several enzymes the RNA is extracted and converted to DNA and with PCR many copies are made.
Now, the DNA bases are made to pair with another bases tagged with markers. Further, the pairs are examined through fluorescence technique. If there are abnormalities, in the pairs the pairing does not occur. In other words, the person stated positive in the test.
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