The term “real-time reverse transcription polymerase chain reaction test” (often abbreviated as “RT-PCR Test”) describes a type of analysis. The real-time polymerase chain reaction (RT-PCR) test technique is utilized for identifying nucleic acid in SARS-CoV-2 in nasopharyngeal or oropharyngeal swab specimens from individuals who have been diagnosed with COVID-19. In order to do this, a process named quantitative PCR, which makes use of fluorescence to monitor the amplification reaction is implemented (qPCR).
Professionals are able to conduct a quick review of the results thanks to this. The reverse transcription-polymerase chain reaction, or RTPCR test in Delhi, is among the procedures that are used rather frequently for identifying COVID-19.
Here are the detailed steps for the RT-PCR test in Delhi
1. Sample collection
A swab will be used to gather any respiratory debris that is detected in your nose when you visit a medical practitioner. A swab has a round, pliable tip that is attached to a long, flexible stick and is designed to be inserted into the nasal cavity. There are a few distinct varieties of nasal swabs, the most common of which are nasal swabs & nasopharyngeal swabs. Nasal swabs gather a sample from just inside your nostrils, while nasopharyngeal swabs collect samples from deeper into the nasal passages. The COVID-19 RTPCR test in Delhi only requires a small amount of sample, so you can collect it with either sort of swab. Following collection, the swab is placed in a tube, where it is then hermetically sealed before being delivered to a laboratory.
2. Sample preparations
For the majority of RTPCR tests in Delhi, either whole RNA or poly(A+) RNA can be employed instead of the other. When working with RNA, it is essential to eliminate all RNases from your solutions, consumables, as well as laboratory equipment. This is one of the most important considerations. You can either buy solutions that are ready to use and free of RNase, or you can treat your own solutions using diethylpyrocarbonate (DEPC) and afterward autoclave the mixture. Both the DEPC treatment and baking at 250 degrees Celsius for three hours are effective methods for inactivating RNases that may be present on laboratory equipment.
3. Reverse Transcription
DNase treatment might be required for prepared RNA samples in order to avoid the possible multiplication of any contaminating genomic DNA. If this were to occur, it could result in an overestimation of the genome size of an mRNA.
A variant of the polymerase chain reaction (PCR) is known as reverse transcriptase–polymerase chain reaction (RT–PCR). The mechanism that is used by both methods is the same, with the exception that the RT–PCR method includes the additional step of performing a reverse transcription of RNA to DNA, also known as RT, in order to facilitate amplification. This indicates that polymerase chain reaction (PCR) is used for pathogens, including such viruses and bacteria which already contain DNA for amplification, whereas reverse transcription polymerase chain reaction (RT–PCR) is employed for pathogens that encompass RNA that requires to be transcribed to DNA in order to achieve the desired result (amplification).
Both of these methods can be carried out in what is known as “real-time,” which indicates that the results can be observed almost as soon as they are produced, in contrast to the “traditional” method, which means that the outcomes can be observed only after the reaction has been completed.
In general, the RTPCR test in Delhi has shown to be the most successful method for determining whether or not the coronavirus is present in the human body. It not only assists in the process of determining whether or not the coronavirus is present, but it also functions as a barrier, preventing the further transmission of infection through contact with an infected individual. The RTPCR test in Delhi from laboratories has been an essential lifesaver throughout the protracted conflict with covid-19.