COVID TEST IN RT-PCR

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. 1 A Seminar as a Part of Curricular Requirement For 1 st Year M. Pharm. 1 st 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. 2 Introduction RT-PCR Genetic material of virus Key steps of RT-PCR Graphical indication Reference CONTENTS

3. 3 As the corona virus that causes the covid19 disease spreads across the world. 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. one of the most accurate laboratory methods for detecting tracking and studying the covid19. INTRODUCTION

4. 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. 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. 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. 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. 8 RT-PCR Work with the covid 19 virus: There are three key steps to the COVID-19 PCR Test Sample collection Extraction PCR sample collection is done using a swab to collect respiratory material found in your nose 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 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 If the virus is present in a sample , these fragments attach themselves to target section of viral DNA Key Steps of RT-PCR

9. 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. 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. 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

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