References materiālu izstrāde un pielietošana SARS-CoV-2 vīrusa noteikšanai klīniskajos paraugos ar reālā laika RT-PCR metodi
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Date
2021
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Rīgas Stradiņa universitāte
Rīga Stradiņš University
Rīga Stradiņš University
Abstract
Pētnieciskā darba “References materiālu izstrāde un pielietošana SARS-CoV-2 vīrusa noteikšanai klīniskajos paraugos ar reālā laika RT-PCR metodi” mērķis bija izstrādāt SARS-CoV-2 references materiālus un pielietot tos reālā laika reversās transkripcijas-polimerāzes ķēdes reakcijas (RT-PCR) metodes izveidei, kas paredzēta vīrusa detektēšanai un tā slodzes noteikšanai siekalu paraugos.
Darbā apkopota literatūra par SARS-CoV-2 vīrusa noteikšanas iespējām un raksturota reālā laika RT-PCR metodes izmantošana vīrusa izraisītās saslimšanas (COVID-19) diagnostikā.
Pētījuma laikā ar molekulārās klonēšanas paņēmienu ieguva vīrusa E gēna fragmentu saturošas plazmīdas, ko izmantoja reālā laika RT-PCR metodes izstrādē. Reālā laika RT-PCR metodes veiktspēju pārbaudīja, analizējot SARS-CoV-2 pozitīvu pacientu nazofaringeālo iztriepju paraugus (n=5). Izstrādāto metodi pielietoja SARS-CoV-2 pozitīvu pacientu siekalu paraugu analīzei (n=42).
The study “Development and application of SARS-CoV-2 reference materials in virus detection in clinical samples by real-time RT-PCR” aimed to create SARS-CoV-2 reference materials and to apply them to establish a real-time reverse transcription-polymerase chain reaction (RT-PCR) based method for virus detection and quantification in saliva samples. The literature review summarized the publications on existing methods of SARS-CoV-2 virus testing and characterised the application possibilities of the real-time RT-PCR method in COVID-19 diagnostics. In this study, a molecular cloning technique was employed to obtain SARS-CoV-2 E gene plasmids used to develop the real-time RT-PCR method. The performance of the method was evaluated by analysing SARS-CoV-2 positive patient nasopharyngeal swabs (n=5). Then, the developed method was applied for saliva sample (n=42) analysis from SARS-CoV-2 positive patients.
The study “Development and application of SARS-CoV-2 reference materials in virus detection in clinical samples by real-time RT-PCR” aimed to create SARS-CoV-2 reference materials and to apply them to establish a real-time reverse transcription-polymerase chain reaction (RT-PCR) based method for virus detection and quantification in saliva samples. The literature review summarized the publications on existing methods of SARS-CoV-2 virus testing and characterised the application possibilities of the real-time RT-PCR method in COVID-19 diagnostics. In this study, a molecular cloning technique was employed to obtain SARS-CoV-2 E gene plasmids used to develop the real-time RT-PCR method. The performance of the method was evaluated by analysing SARS-CoV-2 positive patient nasopharyngeal swabs (n=5). Then, the developed method was applied for saliva sample (n=42) analysis from SARS-CoV-2 positive patients.
Description
Farmācija
Pharmacy
Veselības aprūpe
Health Care
Pharmacy
Veselības aprūpe
Health Care
Keywords
COVID-19, SARS-CoV-2, COVID-19 diagnostika, RT-PCR metode, E gēns, plazmīdas., COVID-19, SARS-CoV-2, COVID-19 diagnostics, RT-PCR method, E gene, plasmids.