Volume 31, Issue 206 (3-2022)
J Mazandaran Univ Med Sci 2022, 31(206): 154-159 |
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Mardomi A, abediankenari S. Introducing a New SYBR green Real-time PCR for Detection of SARS-CoV2 Virus Genome. J Mazandaran Univ Med Sci 2022; 31 (206) :154-159
URL: http://jmums.mazums.ac.ir/article-1-17471-en.html
URL: http://jmums.mazums.ac.ir/article-1-17471-en.html
Abstract: (1490 Views)
Background and purpose: There are various methods for molecular detection of SARS-CoV2 genome among which, PCR-based methods are the most reliable for making diagnosis. The majority of approved PCR kits for detection of Coronavirus are based on TaqMan real-time PCR which is expensive due to incorporating fluorescent and quencher-harboring probe. The aim of this study was to design a simple and affordable method for detection of SARS-CoV2 based on a more affordable SYBR green qPCR method.
Materials and methods: Specific primers were designed for the virus nucleocapsid gene and the human control gene using Oligo software. The specificities of the primers were examined by Primer-BLAST capability according to NCBI database. Nasal swab samples were obtained from 10 PCR-confirmed COVID-19 patients and 10 healthy control people.
Results: The designed reactions were performed in full compliance with the kit approved by the Ministry of Health to discriminate healthy individuals from COVID-19 patients. The Ct cut-off values calculated for N1 and N2 reactions were 39.72 and 39.69, respectively.
Conclusion: The designed SYBR green-based reaction showed a potential role for detection of SARS-CoV2 genome in clinical samples and this method can be considered as a less-expensive alternative to TaqMan real time PCR if the primers and standard reaction conditions are properly designed.
Materials and methods: Specific primers were designed for the virus nucleocapsid gene and the human control gene using Oligo software. The specificities of the primers were examined by Primer-BLAST capability according to NCBI database. Nasal swab samples were obtained from 10 PCR-confirmed COVID-19 patients and 10 healthy control people.
Results: The designed reactions were performed in full compliance with the kit approved by the Ministry of Health to discriminate healthy individuals from COVID-19 patients. The Ct cut-off values calculated for N1 and N2 reactions were 39.72 and 39.69, respectively.
Conclusion: The designed SYBR green-based reaction showed a potential role for detection of SARS-CoV2 genome in clinical samples and this method can be considered as a less-expensive alternative to TaqMan real time PCR if the primers and standard reaction conditions are properly designed.
Type of Study: Brief Report |
Subject:
Biotechnology
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