How Real Time RT-PCR Helps In Detecting COVID-19 Virus?

While COVID-19 continues its dominance all over the world, FAO (Food and Agriculture Organization of the United Nations) in collaboration with IAEA is going to offer its support to incorporate other subcontinent to adopt real time RT-PCR.

It is the most efficient and accurate method to detect, study and track the presence of the Corona virus (COVID-19) in the human body. Are you new to the term real time RT-PCR? Excited to know more about it and its differentiation from that of PCR?

Then, surely you have to go through this blog once as it entails down some fresh details on genetics and viruses as well. So, can we start?

What is actually real time RT-PCR?

Real time RT-PCR test for detecting COVID-19 is a method derived from nuclear reaction to spot special genetic materials in the pathogens (mainly virus). Earlier, it was only used for detecting the targeted genetic materials by radioactiveisotope marketers. But later isotopic labelling is substituted by subsequent refining fluorescent dyes.

Although scientists are able to derive the result instantly yet the process is still under developmental process. But on the other hand, RT-PCR offers the result as soon as it ends! At present real time RT-PCR gets used in laboratories to spot out the presence of COVID-19 virus from the collected samples. 

Previously, it was used to look for the trace of Zika virus and Ebola! But now it should be adapted by every country to detect the COVID-19 virus in real time by increasing its overall testing abilities throughout the world.  

What are this virus and its genetic material?

Virus is actually microscopic organism containing genetic material in a specific molecular envelope. But the genetic material can contain either RNA (ribonucleic acid) or DNA (deoxyribonucleic acid). 

RNA is single-stranded molecule which transmits, transcribes and multiplies its genetic code and proteins. They do so for synthesizing and continuing the performance to sustain the development of the organisms and its breed. Varied RNAs are there which trigger transmission, transcription and multiplication. 

DNA is double-stranded molecule present in every organism including microorganisms, plants and animals. It is the blueprint of the developmental structure of the individual organism.

Virus like SARS-CoV-2 which is responsible for emerging the disease COVID-19 only features RNA. It depends on the healthy cells infiltration for its own multiplication and sustainability in the body. It makes use of its specific genetic code or RNA into the cell for reprogramming of the cell enabling them to act as virus-generating factories.

So, the detection of COVID-19 is only possible by making use of RT-PCR test as it assists in RNA conversion into DNA. The procedure involves in doing so is termed as RT or reverse transcription. It is reliable for multiplying DNA to figure out the presence of the virus in the collected sample.

Amplification is the pivotal process of this entire procedure to multiply the transcribed viral DNA strands into thousands of number. Rather than spotting the minuscule of the virus amount, it helps in detecting large amount of virus for confirmation the infection in the patient’s body. 

How real time RT-PCR performs to detect COVID-19 virus?

To perform RT-PCR test for figuring out the trace of COVID-19 in the body, saw sample is accumulated from the throat or nose of the person. Then it is immersed in numerous chemical solutions for deposition of the fats and proteins for the extraction of RNA strains only. But in this case, RNA is the mixture of the virus’s RNA and genetic material of the individual.

Then reverse transcription (RT) is used so that RNA gets converted into DNA by special enzyme treatment. Those tiny DNA fragments are also complemented with viral DNA which has to be figured out. If virus is there then it will attack the healthy DNA fragments resulting in the genetic fragments to develop DNA strands while others are just left over to check whether it invades on them or not!

At last the strands are positioned in the RT-PCR machine. Then it is cycled via random cooling and heating so that the mixture gets triggered to respond certain chemical reactions to develop identical versions yet new viral DNA target sections in a large amount. Every cycle doubles the present number of viral DNA.

If any RT-PCR is set to perform 35 cycles then it will end up with 35 million of new viral DNA from a little amount of viral sample. On completion of the procedure the viral is labelled and released under fluorescent dye. After certain amount of the dye surpasses through it the presence of the virus will get confirmed.

If less amount of dye needs to reach this level then the infection is severe, otherwise it is confirmed as acute and possesses a high chance to get cleared after 14 days. 

Why using real time RT-PCR tests to detect COVID-19 virus?

The technique of real time RT-PCR is really specific and sensitive which is why it is able to serve you with a reliable diagnostic within 3 hours. But most of the testing laboratories consume 6-8 hours or even a couple of days to deliver the test report due to its high demand and consistent analyse of the collected samples.

In contrast to other virus detection methods, it can be performed quicker and possesses little chance to offer contamination error. Another reason for this it is performed completely in an enclosed tube and regarded as the accurate method to detect COVID-19 so far.

But it is not reliable to detect the past infection that is equally important for determining the transmission and development of this gnarly viral infection. It is significant because this virus invades the human body for a specific window. That’s why other methods are equally vital for studying, tracking and detecting the past infections especially that don’t reveal any symptoms or signs in the body.  

How real time RT-PCR is different from PCR?

Basically, RT-PCR is one of the kinds of PCR (Polymerase Chain Reaction). Both of the methods make use of similar technique. The only differentiation between them is the former one prefers an additional method- reverse transcription (RT) of RNA to DNA.

Actually, it influences the occurrence of amplification and it clearly states that PCR is preferable for using DNA loaded bacteria and viruses. On another flip, RT-PCR is highly preferable for RNA-equipped viruses where RNA transcription or amplification is needed to convert it into DNA.

Both of these are real-time and can deliver accurate result in no time if done in appropriate manner. PCR is preferred as diagnostic test to detect pathogens of the viruses causing foot-and-mouth disease, African swine fever and Ebola. As COVID-19 contains RNA only, RT-PCR is the only method to detect its presence in the human body. 

These are all you should know about real time RT-PCR test and Corona virus! If you are suspecting the virus exposure and want to get next-day results for PCR testing in London then you can get in touch with us! You are always welcome!