AUTHOR=Gao Junshan , Zhang Le , Xue Liang , Cai Weicheng , Qin Zhiwei , Yang Jiale , Liang Yanhui , Wang Linping , Chen Moutong , Ye Qinghua , Li Ying , Wang Juan , Wu Shi , Wu Qingping , Zhang Jumei 

TITLE=Development of a High-Efficiency Immunomagnetic Enrichment Method for Detection of Human Norovirus via PAMAM Dendrimer/SA-Biotin Mediated Cascade-Amplification

JOURNAL=Frontiers in Microbiology

VOLUME=Volume 12 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.673872

DOI=10.3389/fmicb.2021.673872

ISSN=1664-302X

ABSTRACT=Human norovirus is a common cause of acute gastroenteritis worldwide, and oysters have been found to be the main carriers for its spread. The lack of efficient pretreatment methods has been a major bottleneck limiting the detection of viruses in oysters. In this study, we established a novel immunomagnetic enrichment method using polyamidoamine (PAMAM) dendrimer/SA-biotin-mediated cascade amplification for for reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) detection. We compared the capture efficiency of traditional immunomagnetic enrichment, biotin-amplified immunomagnetic enrichment, and PAMAM dendrimer/SA-biotin-mediated cascade-amplification immunomagnetic enrichment. The optimal capture efficiency of the novel method was 44.26% ± 1.45%, which increased by 183.17% and 18.09% compared with the first two methods, respectively. The three methods were used to detect norovirus in 44 retail oysters. The detection rate of the PAMAM dendrimer/SA-biotin-mediated method was 25.0%, which was higher than that of the other two methods by 6.2% and 9.1%, respectively. This study aimed to reduce the economic cost, time, and increase the detection rate. This method can be applied for the rapid detection of norovirus in oysters.