Everything You Need To Know About RT-PCR Testing

What is an RT-PCR test?

The COVID-19 RT-PCR test is a Real-time reverse transcription polymerase chain reaction test for the detection of nucleic acid of SARS-CoV-2 by taking nasopharyngeal or oropharyngeal swab samples from people with suspected COVID-19.

This is achieved by monitoring the amplification reaction using fluorescence, a technique called quantitative PCR (qPCR). It helps professionals review the results right away. RT-PCR is one of the most widely used methods for detecting COVID-19.

RT-PCR is a laboratory technique that involves reverse transcribing (RT) viral RNA into DNA, and then that complementary DNA is amplified using another technique called the polymerase chain reaction (PCR). It helps to detect even a very low virus concentration at this point in time by amplifying the DNA, even if the symptoms of the disease are not visible.

How does a Covid-19 RT-PCR test work?

Sample collection is performed with a swab to collect respiratory material found in the nose. The RT-PCR test uses nasopharyngeal swabs / oropharyngeal swabs / throat swabs / nasal swabs to identify virus particles in patients. The test is carried out by trained specialists using an RT-PCR test kit.

Once removed, the swab is sealed in a tube and then sent to a Covid-19 testing laboratory. It still includes the extraction of genetic material (RNA) from the Covid-19 virus.

The PCR step then uses PCR machine, a so-called thermal cycler that amplifies the genetic material of the SARS-CoV-2 virus. During this process, one of the probes / chemicals will generate a fluorescent light if SARS-CoV-2 is present in the sample.

Who Should Get the RT-PCR Test?

If the patient experiences any of the symptoms of the coronavirus, which is primarily a loss of smell or taste, and difficulty breathing, it is advisable to have the Covid-19 RT-PCR test performed according to ICMR guidelines.

The only way to counter the spread of the pandemic is to conduct rampant testing, and some companies like SpiceHealth have been constantly innovating to provide medical care.

How Accurate are RT-PCR Tests?

RT-PCR is considered to be the Gold standard to detect COVID-19. It's one of the fastest and most accurate laboratory methods for detecting COVID-19. Real-time RT-PCR technology is highly sensitive and specific and can provide a reliable diagnosis in around three to four hours.

According to health experts, RT-PCR tests are more accurate when compared to other tests such as antigen and antibody tests. The positive test result is highly confirmatory, as a double test is carried out before reporting, first during the screening and then for confirmation. Likewise, There are no side effects of the RT-PCR test.

What are the RT-PCR test protocols?

The quantification of mRNA with RT-PCR can be done as follows: a step or a two steps Reaction. The difference between the two approaches lies in the Number of tubes used in carrying out the procedure.

In a one-step protocol, the (RT) reverse transcription and PCR components are mixed simultaneously in a single tube and buffer. Since both reactions take place in the same tube and fewer pipetting steps reduce the risk of contamination, there are relatively fewer experimental variations. It is useful for reinforcing targets that are relatively abundant.

In two-step protocols, the reverse transcription (RT) and PCR steps are carried out in separate tubes and reaction conditions.

A stable pool of complementary DNA (cDNA) is generated here, which can be stored for long periods of time. In addition, the genes from the same cDNA pool can be amplified without multiplexing. Thanks to its highly flexible priming option with various optimized buffers, it achieves significantly better results.

How much time does it take to generate RT-PCR results?

Many test centers process a large number of tests on a daily basis, which delays the results. As a result, certain companies or agencies have sprung up trying to make these services available more quickly.

For example in SpiceHealth Covid-19 test laboratories. The test results will be provided within 24 hours of the specimen collection.

What do the results of the Covid-19 RT-PCR test mean?

The SARS-CoV2-RNA can usually be detected in nasopharyngeal / oropharyngeal smears in the acute phase of infection. positive The results indicate an active infection. If your test is positive, take whatever steps are necessary, see a doctor, stay home, and isolate yourself from others.

negative The results indicate that no RNA specific for SARS-COV-2 is detected. If your test is negative, continue to take preventative measures to protect yourself from contracting COVID-19.

If clinically suspected, a retest is recommended if the results Inconclusive. A non-diagnostic tool Result means that no internal control is recognized, samples must be repeated.

Does a negative PCR result certify a full recovery from Covid-19?

A negative PCR test result simply means that the person is not currently affected by Covid-19. It neither confirms recovery from COVID-19 nor does it confirm that the patient is no longer infectious.

About the author:

Dinesh Mittal is a freelance content marketer with over 4 years of experience curating content and posts in the fields of health, psychology, mental health and mental wellbeing.