How is the corona virus detected using RT-PCR(reverse transcriptase polymerase chain reaction)?

Different countries used different protocols for RT-PCR diagnostic testing for SARS CoV-2. Some countries target the N region gene of SARS CoV-2. This is because, among other reasons, it is a highly conserved region in coronaviruses genome and to deliver consistent results. so basically we need to target specific genes and perform RT-PCR test.

All sequence of SARS CoV-2 first provided by china in december 2019. https://www.ncbi.nlm.nih.gov/nuccore/NC_045512

genome sequence of SARS CoV-2(coronavirus)
SARS CoV-2 genome. Orf (open reading frame), S (spike gene), E(envelope gene, M (membrane gene), N(nucleocapsid region)

METHOD of RT PCR

First take sample through nasopharyngeal or oropharyngeal swap. Once swap is take then immediately transfer into viral transport medium(to protect viral genome). Then extract RNA using buffer solution.Conventionally in PCR DNA is used for amplification process so first convert RNA into DNA by using reverse transcriptase.

Then perform PCR followed by three steps

  • DENATURATION:- separate the DNA double helix
  • ANNEALING:- Primers binds the specific region(N-region) and starts polymerisation
  • EXTENSION:- New strands of DNA are made using original strand are templates DNA polymerase enzyme joins free DNA nucleotide together.

PCR is cycle process repeated many times in 2-3 hours generate billion of copies.

To get result we need to know PCR annealing step in detail

Primers are small fragments of ssDNA designed to bind specifically to a single region in the genome to allow for precise amplification of the diagnostic target area. Primers bind to the target area and give off a flurescent signal as amplification of the area increases. This flurescent signal is then read by the quantitative real PCR machine where the reaction is occuring to give a result positive or negative.

If sample test positive covid-19
IF SAMPLE TEST NEGATIVE FROM COVID-19

In india COVID-19 test conducted as per kits approved by ICMR(indain council of medical research)

manoj kumar

Manoj kumar singh is a digital marketing expert and content writer at Pharmshala .

5 thoughts on “How is the corona virus detected using RT-PCR(reverse transcriptase polymerase chain reaction)?

  • June 29, 2020 at 12:20 pm
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    Great initiative bhaii, keep the spirit up…🤩🔥✨

    • June 29, 2020 at 12:59 pm
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      thanks

  • June 29, 2020 at 12:28 pm
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    Thanks for this information

    • June 29, 2020 at 12:59 pm
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      thanks

  • June 29, 2020 at 3:15 pm
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    So did we get any results?? By experimenting on N region of genome of virus?

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