Instrumental techniques in detecting COVID-19 infection

Cilj: Opisati instrumentalne tehnike i objasniti njihovu primjenjivost u otkrivanju COVID-19 infekcije. Materijal i metode: Opisan je postupak izvođenja RT-qPCR reakcije od izolacije RNA do detekcije kao i RT-LAMP metode. Prikladni materijal potrebni za postupke RT-qPCR-a i RT-LAMP-a su također opisani. Rezultati: Jačina fluorescencije PCR amplikona se predočava elektroferogramom na kojem se prikazuje na apscisi broj ciklusa reakcije, a na ordinati jačina fluorescencije. Ct vrijednost je obrnuto proporcionalna količini virusa mjerenog uzorka. Za RT-LAMP metodu će se na ekranu aparata pokazati pozitivan, negativan ili neispravan rezultat. Zaključak: RT-qPCR je zlatni standard u detekciji COVID-19 infekcija zbog svoje visoke preciznosti i osjetljivosti. RT-LAMP će se u bliskoj budućnosti više koristiti zbog brzine detekcije i jednostavnosti uporabe. Sekvenciranje je ključ u identificiranju različitih podtipova SARS-CoV-2.Objectives: To describe instrumental techniques and explain their applicability in the detection of COVID-19 infection. Material and methods: The procedure for performing the RT-qPCR reaction from RNA isolation to detection is described, as well as the RT-LAMP method. Appropriate material required for RT-qPCR and RT-LAMP procedures are also described. Results: The fluorescence intensity of the PCR amplicon is represented by an electropherogram showing the number of reaction cycles on the abscissa and fluorescence intensity on the ordinate. The Ct value is inversely proportional to the amount of virus in the measured sample. For the RT-LAMP method, a positive, negative or invalid result will be displayed on the device screen. Conclusion: RT-qPCR is the gold standard in the detection of COVID-19 infections due to its high precision and sensitivity. RT-LAMP will be used more in the near future due to its speed of detection and ease of use. Sequencing is the key to identifying the different subtypes of SARS-CoV-2