BIOEDUSCIENCE
Vol 6 No 2 (2022): BIOEDUSCIENCE

Optimasi Volume Kit Da An Gene Untuk Deteksi SARS-CoV-2 dengan Real Time RT-PCR

Seprianto (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)
Muhammad Arreza (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)
Titta Novianti (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)
Febriana Dwi Wahyuni (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)
Oktaviani Naulita Turnip (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)
Roaslein Putri (PT. Ecosains Hayati, Jakarta, Indonesia 13550)
Henny Saraswati (Program Studi Bioteknologi, Fakultas Ilmu-ilmu Kesehatan, Universitas Esa Unggul, Jakarta, Indonesia 11510)



Article Info

Publish Date
31 Aug 2022

Abstract

Background: SARS-CoV-2 is a new type of coronavirus of the genus Betacoronavirus and the family Coronaviridae that causes a respiratory disease called COVID-19. The virus has a sheath and genetic material in the form of single-chain RNA. The genome structure of this virus is divided into two types, namely genes that encode non-structural proteins consisting of the ORF1a / ORF1b gene and genes that encode structural proteins consisting of spike glycoprotein (S), envelope (E), membrane glycoprotein (M), and nucleocapsid protein (N). Methods: The method of detecting SARS-CoV-2 with real time RT-PCR is the most recommended method because it has high specificity and accuracy. The specificity of a method is necessary to be able to specifically recognize the pathogen that causes the disease. Real time RT-PCR requires sampling with a swab on the oropharynx or nasopharynx to be examined in the laboratory which later the presence of viral RNA becomes a molecule that is assessed for diagnosis results. In this study, volume optimization was carried out on the Da An Gene kit used for the detection of SARS-CoV-2 with Reverse Transcription Polymerase Chain Reaction (Real time RT-PCR) with the aim of saving the use of reagents from available kits but with amplification results remaining optimal and accurate. Results: There were three SARS-CoV-2 RNA samples used consisting of N62, N63, and N79 samples and three types of total volume used were 20 μl, 15 μl, and 10 μl. The results of this study showed that the three positive samples contained SARS-CoV-2 with a Cq value of < 40. Conclusion: A volume of 20 μl is the optimal volume, which is more efficient than the manufacturer's recommended volume of 25 ul.

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Journal Info

Abbrev

bioeduscience

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education

Description

BIOEDUSCIENCE is an open access journal that publishes research in the field of Biology and Biosain Education such as: Applied and implemented in education and learning, Botany, Zoology, Microbiology, Ecology, Biotechnology, Molecular Biology, Genetics, Bioinformatics, Cell and Developmental ...