Name: Juhany M.

Musa

Section: BSMT 1-B

Coronavirus Test: Real time RT-PCR

Description of the Assay:

Real time RT-PCR (Reverse Transcription Polymerase Chain

Reaction) is a diagnostic test used to detect the presence of viral genetic
material (RNA) from the coronavirus, which causes COVID-19. This test is
considered the gold standard for COVID-19 diagnosis.

The test involves collecting a sample from a patient's respiratory tract using a
swab or other collection device. The collected sample is then sent to a
laboratory where it undergoes a complex series of processes to extract RNA
from the virus.

Once the RNA is extracted, it is converted into DNA using an enzyme called
reverse transcriptase. The resulting DNA is then amplified using a technique
called polymerase chain reaction (PCR), which produces millions of copies of
the viral DNA.

During the PCR amplification process, fluorescent probes are added to the
reaction mixture that binds specifically to the viral DNA. The amount of
fluorescent signal generated is measured in real-time, which allows for the
quantification of the viral load in the original sample.

The test results are usually available within a few hours, and a positive result
indicates the presence of the coronavirus in the patient's sample. This test is
highly sensitive and specific, meaning it can accurately detect the virus even at
low levels of infection.

How it works:

Real-time RT-PCR (Reverse Transcription Polymerase Chain

Reaction) works by detecting the genetic material (RNA) of the coronavirus in
a patient's sample. The test involves several steps:
 1. Sample Collection: A sample is collected from the patient's respiratory
tract, usually through a nasal or throat swab.
2. RNA Extraction: The genetic material (RNA) of the virus is extracted
from the sample in a laboratory. This is done by breaking open the
virus particles and separating the RNA from other materials in the
sample.
3. Reverse Transcription: The RNA is then converted into
complementary DNA (cDNA) using a special enzyme called reverse
transcriptase. This step allows the RNA to be detected by the PCR
reaction.

4. PCR Amplification: The cDNA is then amplified using a technique

called polymerase chain reaction (PCR). This involves heating and
cooling the reaction mixture to create millions of copies of the cDNA.

5. Detection: During the PCR amplification process, fluorescent probes

are added to the reaction mixture that binds specifically to the viral
DNA. The amount of fluorescent signal generated is measured in real-
time using specialized instruments, which allows for the quantification
of the viral load in the original sample.

If the patient's sample contains the coronavirus, the test will detect the viral
RNA and generate a positive result. If the sample does not contain the virus,
the test will produce a negative result.

Real-time RT-PCR is a highly sensitive and specific test that can detect the
virus even at low levels of infection. It is considered the gold standard for
COVID-19 diagnosis.
Major Breakthrough:

Real-time RT-PCR (Reverse Transcription Polymerase Chain

Reaction) is considered a major breakthrough in the diagnosis of COVID-19.
This diagnostic test has revolutionized the way the virus is detected and has
played a crucial role in controlling the spread of the disease.

Before the development of real-time RT-PCR, the detection of the coronavirus

was a complex and time-consuming process that required several days to
obtain the results. The development of this test has greatly reduced the time
required for diagnosis, with results typically available within a few hours.

Real-time RT-PCR is highly sensitive and specific, meaning it can detect the
virus even at low levels of infection. This has enabled early detection of the
virus and facilitated the implementation of appropriate public health measures
such as contact tracing and isolation to prevent the spread of the disease.

Furthermore, real-time RT-PCR has enabled researchers to study the virus in

greater detail, including its transmission patterns, genetic mutations, and the
effectiveness of vaccines and treatments. This information has been critical in
developing effective strategies to control the spread of the disease and save
lives.

Overall, the development of real-time RT-PCR has been a major breakthrough

in the fight against COVID-19, and its continued use is essential in controlling
the pandemic.
Closing remarks:

Real-time RT-PCR has played a crucial role in the diagnosis of

COVID-19, allowing for early detection and control of the virus. Its high
sensitivity and specificity have enabled public health measures such as contact
tracing and isolation, helping to slow the spread of the disease. Furthermore,
this diagnostic test has enabled researchers to gain a better understanding of
the virus, facilitating the development of effective treatments and vaccines.

As the COVID-19 pandemic continues, real-time RT-PCR remains a critical

tool in the fight against the virus. It is important to continue to prioritize
testing efforts and ensure that diagnostic resources are available and accessible
to those who need them. By working together and utilizing the latest
diagnostic technologies, we can continue to make progress in controlling the
spread of the virus and ultimately bring an end to the pandemic.