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KMID : 1144820210270040283
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대한의생명과학회지
2021 Volume.27 No. 4 p.283 ~ p.290
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Development of Reverse Transcriptase Polymerase Chain Reaction Primer Sets and Standard Positive Control Capable of Verifying False Positive for the Detection of Severe acute respiratory syndrome coronavirus 2
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Cho Kyu-Bong
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Abstract
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Severe acute respiratory syndrome coronavirus (SARS-CoV2) is a major coronavirus that infects humans with human Coronavirus (HuCoV)-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL63, Severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle east respiratory syndrome coronavirus (MERS-CoV). SARS-CoV2 is currently a global pandemic pathogen. In this study, we developed conventional RT-PCR based diagnostic system for the detection of SARS-CoV2 which is relatively inexpensive but has high stability and a wide range of users. Three conventional RT-PCR primer sets capable of forming specific band sizes by targeting the ORF1ab [232 nucleotide (nt)], E (200 nt) and N (288 nt) genes of SARS-CoV2 were developed, respectively, and it were confirmed to be about 10~100 times higher detection sensitivity than the previously reported methods. In addition, a standard positive control that can generate specific amplicons by reacting with the developed RT-PCR primers and verify the false-positiv from contamination of the laboratory was produced. Therefore, the diagnostic system that uses the RT-PCR method is expected to be used to detect SARS-CoV2.
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KEYWORD
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Conventional RT-PCR, SARS-CoV2, Severe acute respiratory syndrome coronavirus, Standard positive control
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