RT-PCR test

Understanding the COVID-19 RT-PCR test

The year 2020 has left no stones unturned to be the worst year of the century. The Covid-19 being the major of all the nightmares humanity has ever had. It took the lives of more than 134,804 humans till now. Moreover above this, there is still a long wait for the vaccine. In contrast to this, we humans have become successful in adopting activities such as practicing yoga/pranayam, preferring Ayurvedic medication over allopathy, eating home-cooked veg meal, and other immunity-boosting ailments.

There is no problem in saying that many have overcome the fear of the coronavirus by doing breathing exercises. But many have also underestimated this corona nightmare. Soon or later they pay the price for it. Hence it goes without saying that till a cure is not out against the virus, our masks are the most effective vaccines. 

There are varied precautionary treatments developed since the beginning of the spread of the virus. The COVID-19 RT-PCR test also came out which you must have heard by now. As the number of coronavirus positive cases in several parts of India witnessed a hike, Maharashtra has made it mandatory for all travelers arriving in the state from Delhi, Gujarat, Goa, and Rajasthan to carry a Real-Time Polymerase Chain Reaction (RT-PCR) negative test report for COVID-19. To get a brief review of the test keep reading.

What is RT-PCR Test?

The COVID-19 RT-PCR test is a real-time reverse transcription-polymerase chain reaction (rRT-PCR) test. The test is for the qualitative detection of nucleic acid from SARS-CoV-2 in upper and lower respiratory specimens (such as nasopharyngeal or oropharyngeal swabs) collected from individuals suspected of COVID-19.

Reverse transcription-polymerase chain reaction is a laboratory method combining reversed 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. In India, the rates for RT-PCR test varies state wise. 

How this Test works?

The RT-PCR test works by testing the collected swab from the respiratory tract of the individual/patient to detect the virus’s genetic material. The process involves sticking a 6-inch swab deep into your nose to collect virus samples to test.

The RNA isolated from upper and lower respiratory specimens (such as nasal, nasopharyngeal or oropharyngeal swabs, sputum, lower respiratory tract aspirates, bronchoalveolar lavage, and nasopharyngeal wash/aspirate or nasal aspirate) is reverse transcribed to cDNA and subsequently amplified using Applied Biosystems QuantStudio7 Flex (QS7) instrument. This is the basic principle of the RT-PCR test. Some complex instruments are taken in use during the diagnoses. 

Reagents used during the test

Reagents

DNA and Viral Small Volume Kit (3×192 purifications)

MagMAX Viral/Pathogen Nucleic Acid Isolation Kit

TaqPath™ 1-Step Multiplex Master Mix (No ROX) COVID-19_N1-F Primer

COVID-19_N1-R Primer

COVID-19_N1-P Probe

COVID-19_N2-F Primer

COVID-19_N2-R Primer

COVID-19_N2-P Probe

COVID-19_N3-F Primer

COVID-19_N3-R Primer

COVID-19_N3-P Probe

RP-F Primer IDT

RP-R Primer

RP-P Probe

COVID-19_N_Positive Control

Hs_RPP30_Internal Extraction Control

Who can take RT-PCR Test?
  • All patients with “mild” symptoms.
  • Patients with exposure risks and no symptoms.
  • Whereas individuals with severe symptoms are advised to seek immediate care.
How to take RT-PCR Test?

The test can also be used for nasal swab that is self-collected by the individual using professional testing kits such as 

  • The Pixel by LabCorp COVID-19 test home collection kit by individuals when determined by a healthcare provider (HCP) to be appropriate based on results of a COVID-19 questionnaire
  • The LabCorp At Home COVID-19 test home collection kit when directly ordered by a HCP.
Accuracy of the test

If performed rightly, RT-PCR swab tests would be pretty close to 100 percent accurate. To get the most accurate results, RT-PCR tests should be conducted 8 days after suspected exposure or infection, to ensure that enough viral material is present to detect. In India medical professionals and the Government claim to provide the test results within 6 hours of the sample collection.

State-wise cost of RT-PCR test
State Current cost Previous cost
Maharashtra Rs 980 Rs 4,500
Delhi -Rs 2,400

-Rs 499 (by Spice Health – a subsidiary of SpiceJet airline)

Karnataka Rs 800
Gujarat Rs 1,500 Rs 2,500
Utter Pradesh Rs 600 Rs 1,600
Kerala Rs 2,100 Rs 2,750
Andra Pradesh Rs 1,600 Rs 2,400
West Bengal Rs 1,500 Rs 2,250
Rajasthan Rs 1,200 Rs 2,200
Assam Rs 2,200/Free
Meghalaya Rs 1000
Telangana Rs 850 Rs 2,200
What you need to know before taking the test?
  • Negative results from pooled testing should not be treated as definitive. If a patient’s clinical signs and symptoms are inconsistent with a negative result or results are necessary for patient management, then the patient should be considered for individual testing. 
  • Specimens with low viral loads may not be detected in sample pools due to the decreased sensitivity of pooled testing.
  • Results are for the identification of SARS-CoV-2 RNA. The SARS-CoV-2 RNA is generally detectable in respiratory specimens during the acute phase of infection. 
  • Positive results are indicative of the presence of SARS-CoV-2 RNA; clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. 
  • Positive results do not rule out a bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of the disease.
  • Negative results do not hinder SARS-CoV-2 infection and should not be used as the sole reason for patient supervision decisions. 
  • Negative results must be combined with clinical observations, patient history, and epidemiological information. 
  • COVID-19 RT-PCR testing is intended for use by trained clinical laboratory personnel specifically guided and trained in the procedures of real-time PCR and in vitro diagnostic methods. 
  • The COVID-19 RT-PCR test must be only used under the US-FDA’s Emergency Use Authorization (EUA) protocol.
Warnings
  • This test has not been FDA cleared or approved. 
  • This test has been authorized by the FDA under an Emergency Use Authorization (EUA) only. 
  • This test has been authorized only for the detection of nucleic acid from SARS-CoV-2, not for any other viruses or pathogens.

Check more updates on COVID-19

4 thoughts on “Understanding the COVID-19 RT-PCR test”

  1. What is the likely hood of:
    a. testing covid positive BUT having a different dominant infection? looks like a gamble test to me?
    b. the annealing process alter the test tube crystal structure and in so doing alter the contents of the test tube?
    c. ,at 35 cycles (+34bill amplifications / replication), there being room left in the test tube without damaging existing copies.
    d. industrial scale demands making human error high enough to have a large margin of error in the results.

  2. THE RTCPR TEST HAS “NOT” BEEN APPROVED AND ‘CANNOT’ DIAGNOSE OR TREAT A DISEASE! PLEASE CHECK OUT THIS TRUTH VIDEO REGARDING THIS TEST AND THE “FACT” THAT THIS TEST IS VIOLATING THE FEDERAL TRADE COMMISSION ACT! EVERY TEST DONE BY THIS ‘RTCPR TEST ‘ THAT PRODUCES A POSITIVE RESULT, IS IN VIOLATION OF THE FED. TRADE COMMISSION ACT! THERE ARE PEOPLE SUFFERING AND DYING DUE TO THIS “UNAPPROVED” TEST!
    https://www.youtube.com/watch?v=p_hwJkhNo9w&feature=youtu.be

  3. THE RTCPR TEST HAS “NOT” BEEN APPROVED AND ‘CANNOT’ DIAGNOSE OR TREAT A DISEASE! PLEASE CHECK OUT THIS TRUTH VIDEO REGARDING THIS TEST AND THE “FACT” THAT THIS TEST IS VIOLATING THE FEDERAL TRADE COMMISSION ACT! EVERY TEST DONE BY THIS ‘RTCPR TEST ‘ THAT PRODUCES A POSITIVE RESULT, IS IN VIOLATION OF THE FED. TRADE COMMISSION ACT! THERE ARE PEOPLE SUFFERING AND DYING DUE TO THIS “UNAPPROVED” TEST!
    https://www.youtube.com/watch?v=p_hwJkhNo9w&feature=youtu.be

  4. **IF THE MODERATOR REFUSES TO POST MY COMMENT, IT WILL BE BECAUSE, OF THE FACT THAT WHAT I POSTED IS IN ‘FACT’ THE “TRUTH!”
    THE RTCPR TEST HAS “NOT” BEEN APPROVED AND ‘CANNOT’ DIAGNOSE OR TREAT A DISEASE! PLEASE CHECK OUT THIS TRUTH VIDEO REGARDING THIS TEST AND THE “FACT” THAT THIS TEST IS VIOLATING THE FEDERAL TRADE COMMISSION ACT! EVERY TEST DONE BY THIS ‘RTCPR TEST ‘ THAT PRODUCES A POSITIVE RESULT, IS IN VIOLATION OF THE FED. TRADE COMMISSION ACT! THERE ARE PEOPLE SUFFERING AND DYING DUE TO THIS “UNAPPROVED” TEST!
    https://www.youtube.com/watch?v=p_hwJkhNo9w&feature=youtu.be

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