Extraction of norovirus based on nanomagnetic beads and establishment of a fluorescence quantitative detection method

Abstract

Norovirus infection is the main cause of epidemic acute gastroenteritis outbreaks. As a result, norovirus merits careful attention. Acute gastroenteritis caused by norovirus is very harmful, necessitating effective prevention and monitoring. This study extracted norovirus using the Fe3O4 nanomagnetic bead method while establishing a rapid and sensitive quantitative norovirus detection method for emergency use. Norovirus RNA was extracted from norovirus-positive stool samples, norovirus-specific primers were designed, and SYBR Green I fluorescent dye was used to construct specific fluorescence quantitative reverse-transcription polymerase chain reaction (RT-PCR) method. Norovirus in the sample was successfully detected with the quantitative RT-PCR method established in this study, with a measured cycle threshold value of 28.73; the melting temperature value of the PCR product was approximately 83.5 °C. Additionally, by analyzing melting curve with quantitative PCR and evaluating the repeatability of the detection method, the performance of the reaction system in terms of specificity and repeatability was determined to be good. The fluorescence quantitative detection method established here can rapidly and effectively detect norovirus, and the method has good specificity and high repeatability. Currently, there is a trend of norovirus infection becoming increasingly severe. Thus, this rapid and sensitive fluorescence quantitative detection method will play a significant role in the monitoring and detection of norovirus and is thus worthy of promotion and application.