The development of multiplex one-step real-time rt-pcr for simultaneous detection variants of severe acute respiratory syndrome-coronavirus-2
5
Issued Date
2022
Copyright Date
2022
Resource Type
Language
eng
File Type
application/pdf
No. of Pages/File Size
xi, 72 leaves
Access Rights
open access
Rights
ผลงานนี้เป็นลิขสิทธิ์ของมหาวิทยาลัยมหิดล ขอสงวนไว้สำหรับเพื่อการศึกษาเท่านั้น ต้องอ้างอิงแหล่งที่มา ห้ามดัดแปลงเนื้อหา และห้ามนำไปใช้เพื่อการค้า
Rights Holder(s)
Mahidol University
Bibliographic Citation
Thesis (M.Sc. (Medical and Molecular Microbiology))--Mahidol University, 2022)
Suggested Citation
Natsada Saengdao The development of multiplex one-step real-time rt-pcr for simultaneous detection variants of severe acute respiratory syndrome-coronavirus-2. Thesis (M.Sc. (Medical and Molecular Microbiology))--Mahidol University, 2022). Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/113840
Title
The development of multiplex one-step real-time rt-pcr for simultaneous detection variants of severe acute respiratory syndrome-coronavirus-2
Author(s)
Abstract
This study aimed to develop a multiplex one-step real-time RT-PCR test for the identification of SARS-CoV-2 variants that is quick and less expensive compared to the efficiency of the standard approach, genome sequencing. Moreover, the developed test would be used to investigate the epidemiology of the variants of SARS-CoV-2 in Thailand. Multiplex one-step real-time RT-PCR for variant detection of SARS-CoV-2 was developed with the plasmid positive control, and validated the sensitivity and specificity by using 155 SARS-CoV-2 positive RNA samples. The cross-reactivity of the other viral respiratory pathogen and the limit of detection were evaluated. Moreover, this test was used to investigate the SARS-CoV-2 variants from April 2021 to April 2022 by using 1,200 positive SARS-CoV-2 samples that outbreak each month. The developed test demonstrated that each SARS-CoV-2 variant has a high sensitivity and specificity. There was no cross-reactivity to other respiratory viral pathogens. According to the epidemiology investigation, the primary SARS-CoV-2 outbreak in Thailand between 2021 and 2022 began with the Alpha variation followed by Delta and Omicron (BA.1) and Omicron (BA.2) variants, respectively. The developed multiplex one-step real-time RT-PCR assay described in this study made it possible to simultaneously detect SARS-CoV-2 variants with high sensitivity, and specificity, and help with SARS-CoV-2 surveillance and outbreak. IMPLICATION OF THE THESIS: Suggestions for further research: SARS-CoV-2 generates several novel varieties due to its high rate of mutation. Therefore, to find the new variations and use as a screening method for SARS-CoV-2, a one-step real-time RT-PCR with specific primers and probes could be developed.
Degree Name
Master of Science
Degree Level
Master's degree
Degree Department
Faculty of Medicine Siriraj Hospital
Degree Discipline
Medical and Molecular Microbiology
Degree Grantor(s)
Mahidol University