In this slide contains introduction, genomic materials of virus and testing method of covid 19 by using RT-PCR. Presented by: R.Rekha (Department of pharmacology), RIPER, anantapur.

1. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 1
A Seminar as a Part of Curricular Requirement
For 1st Year M. Pharm. 1st Semester
04-06-2021
Presented by
R.Rekha (20l81S0111)
M.Pharmacy
Department of Pharmacology
Under the guidance of
DR. P. Ramalingam, M.Pharm, Ph.D
Research Director & Professor of Pharmaceutical & Medicinal Chemistry
Covid test in RT-PCR

2. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 2
o Introduction
o RT-PCR
o Genetic material of virus
o Key steps of RT-PCR
o Graphical indication
o Reference
CONTENTS

3. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 3
As the corona virus that causes the covid19 disease spreads across the world.
o The IAEA in partnership with the food and agriculture organization of the united
nations is offering to support and expertise to help countries use real time reverse
transcription polymerase chain reaction.
o one of the most accurate laboratory methods for detecting tracking and studying
the covid19.
INTRODUCTION

4. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 4
Real time RT-PCR is a nuclear- derived method for detecting presence of specific
genetic material in any pathogen including a virus.
Originally, the method used radio active isotopes markers to detect targeted genetic
materials, but subsequent refining has led to the replacement of isotopic labelling
with special markers, most frequently fluorescent dyes RT-PCR only provides
results at the end of process
RT-PCR is one of the most widely used laboratory methods for detecting the covid19
virus
RT-PCR

5. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 5
A virus is a microscopic package of genetic material surrounded by molecular
envelope. This genetic material can be either deoxyribonucleic acid (DNA) or
ribonucleic acid (RNA)
Genetic material of Virus

6. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 6
• DNA is a two strand molecule that is found in all organisms, such as animals,
plants and viruses, and which holds the genetic code, or blueprint, for how
these organisms are made and develop.
• RNA is generally a one-strand molecule that copies, transcribes and transmits
parts of the genetic code to proteins so that they can synthesize and carry out
functions that keep organisms alive and developing. Different variations of
RNA are responsible for copying, transcribing and transmitting.

7. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 7
• Some viruses such as the SARS-CoV-2 carona virus which causes covid-19 only
contain RNA which means that they are healthy cells to multiply and survive
• In order for virus like covid 19 virus to be detected early in the body using real
time RT-PCR this process is called reverse trascription
• DNA can be copied - or amplified – which is a keep art of real time RT-PCR
detected process, hundreds and thousands copies of DNA strands.
• Then the genitic information at target section viral DNA accurately confirm that
the virus is present.

8. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 8
• RT-PCR Work with the covid 19 virus:
There are three key steps to the COVID-19 PCR Test
1) Sample collection
2) Extraction
3) PCR sample collection is done using a swab to collect respiratory material
found in your nose
4) The sample is treated with several chemical solutions that removes
substances such a proteins and fats and that extract only the RNA present
in sample
5) The RNA is reverse transcribed to DNA using a enzyme then add additional
short fragment of DNA that are complementary to specific parts of the
transcribed viral DNA
6) If the virus is present in a sample , these fragments attach themselves to
target section of viral DNA
Key Steps of RT-PCR

9. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 9
• The mixture is then placed in an RT-PCR machine . The machine cycles though a
temperature that heat and cool the mixture though trigger specific chemical
reactions that create new , identical copies of the target sections of viral DNA each
cycles doubles two copies become four, four copies become eight.
• The process around 35 billion new copies of the viral DNA created from each stand
of the virus present in sample.
• DNA strand and then release a fluroscent dye

10. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 10
• The computer tracks the amount of flurosence in the sample after each cycle.
• When a certain level of flurosence is surfaced , this confirms that the virus is
present to estimate the severity of the infection: the fewer the cycles , the more
severe viral infection.
Graphical Indication of virus

11. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11
1.Zhu H, Wei L, Niu P. The novel corona virus outbreak in Wuhan, China. Glob Health
Res Policy.2020;(2) 5:6.
2. Song F, Shi N, Shan F., Zhang Z, Shen J., Lu H. Emerging coronavirus 2019-nCoV
pneumonia. Radiology. 2020;295(1):210-217
3. Faoud T.Ishamael. Principles and applications of polymerase chain reaction.
Elsevier. 2008; 101. (4) : 437- 443.
Reference

12. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 12