Molecular diagnostics of SARS-CoV-2 virus

Cilj rada: Cilj ovog rada bio je teorijski prikazati i razjasniti karakteristike i građu virusa SARS-CoV2, opisati trenutno dostupne dijagnostičke metode za analizu virusa i prikazati najbolju dijagnostičku tehniku, RT-PCR molekularnu metodu. Materijali i metode: U ovom radu opisan je postupak molekularne metode dijagnostike SARS-CoV-2 dijagnostike na BIO-RAD CFX96 RT- PCR uređaju uz korištenje reagensa LiliF Master Mix kit za pripremu reakcijske smjese, koji se koriste u Nastavnomzavodu za javno zdravstvo Splitsko-dalmatinske županije Rezultati: Svi uzorci moraju imati detektiranu IPC (Internu kontrolu) koja dokazuje da je postupak obrade uzorka dobro napravljen, te se može pristupiti interpretaciji nalaza. Pozitivan nalaz na virus SARS-CoV-2 označava prisutnost tri detektibilna gena: E,RdRp i N, dok za negativan rezultat geni nisu eksprimirani. Zaključak: Najbrža, najosjetljivija i najisplativija dijagnostička metoda za detekciju i kvantifikaciju novonastalog virusa jest RT-PCRObjectives: The aim of thisstudy wasto theoretically present and clarify the characteristics and structureof the SARS-CoV-2 virus, to describe the currently available diagnostic methods for the analysis of the virus and to present the best diagnostic technique, the RT-PCR molecular method. Methods: This manuscript describes the procedure of the molecular method for SARS-CoV-2 detection on the BIO-RAD CFX96 RT-PCR instrument using LiliF Master Mix kit reagents for the preparation of the reaction mixture, which are used at the Teaching Public Health Institute of Split-Dalmatia County. Results: All samples must have a detected IPC (Internal Control), which proves that the sample processing procedure was done well, and the interpretation of test results can be accessed. A positive test for SARS-CoV-2 virus indicates the presence of three detectable genes: E, RdRp and N, while for a negative result the genes are not expressed. Conclusion: The fastest, most sensitive and most cost-effective diagnostic method for the detection and quantification of newly emerging SARS-CoV-2 virus is RT-PCR