Journal of Advanced Biomedical Sciences
JABS
Tue, Dec 3, 2024
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Volume 12, Issue 2 (4-2022)
JABS 2022, 12(2): 187-193 |
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Morovvati A, Javadi A, Dehghani sanij S, Zarei M, Molazadeh S, Badakhsh M et al . Rapid Homemade Reverse Transcriptase PCR Detection and Phylogenetic Analysis for SARS-CoV-2 Based on E and M Genes. JABS 2022; 12 (2) :187-193
URL: http://jabs.fums.ac.ir/article-1-2773-en.html
URL: http://jabs.fums.ac.ir/article-1-2773-en.html
Abbas Morovvati1 
, Ali Javadi2 , Somayeh Dehghani sanij1 , Mohamadreza Zarei1 , Shima Molazadeh3 , Mina Badakhsh1 , Mohsen Zargar * 1
, Ali Javadi2 , Somayeh Dehghani sanij1 , Mohamadreza Zarei1 , Shima Molazadeh3 , Mina Badakhsh1 , Mohsen Zargar * 1
1- Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran
2- Department of Pathobiology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, Iran
3- Department of Quality Assurance, Razi Vaccine & Serum Research Institute, Agricultural Research, Karaj, Iran
2- Department of Pathobiology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, Iran
3- Department of Quality Assurance, Razi Vaccine & Serum Research Institute, Agricultural Research, Karaj, Iran
Abstract: (26511 Views)
Background & Objective: Coronavirus disease 2019 spreads worldwide and needs detection systems capable of rapid diagnostic of this virus (SARS-CoV-2). The aim of this study is to design the homemade RT-PCR method for the Detection and phylogenetic analysis of this virus
Material & Methods: The genes selected for diagnosis were E and M genes for this virus. PCR product was cloned in pTZ57R/T plasmid for preparation of positive control. In order to determine the sensitivity of this molecular method, the genes mentioned in the clone pTZ57R/T vector and the Limit of detection (LOD) the genes were determined and phylogenetic analysis was performed using partial E and M gene sequences.
Results: PCR product was observed for E and M genes 156 and 547 bp on the Agarose gel. The LOD of the E and M gene was 60 and 82 copies. There was also a positive response to the samples of patients who were positive by other methods.
Conclusions: Since this virus is considered to be the cause of a pandemic in different countries all over the world, the present study is very important as a method of rapid and low-cost molecular diagnosis for monitoring this virus. Phylogenetic analysis is necessary for epidemiological studies for the control and prevention of the disease.
Material & Methods: The genes selected for diagnosis were E and M genes for this virus. PCR product was cloned in pTZ57R/T plasmid for preparation of positive control. In order to determine the sensitivity of this molecular method, the genes mentioned in the clone pTZ57R/T vector and the Limit of detection (LOD) the genes were determined and phylogenetic analysis was performed using partial E and M gene sequences.
Results: PCR product was observed for E and M genes 156 and 547 bp on the Agarose gel. The LOD of the E and M gene was 60 and 82 copies. There was also a positive response to the samples of patients who were positive by other methods.
Conclusions: Since this virus is considered to be the cause of a pandemic in different countries all over the world, the present study is very important as a method of rapid and low-cost molecular diagnosis for monitoring this virus. Phylogenetic analysis is necessary for epidemiological studies for the control and prevention of the disease.
Type of Study: Research |
Subject:
Virology
Received: 2021/12/25 | Accepted: 2022/03/6 | Published: 2022/06/28
Received: 2021/12/25 | Accepted: 2022/03/6 | Published: 2022/06/28
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This work is licensed under a Creative Commons — Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)