Development of a High-Efficiency Immunomagnetic Enrichment Method for Detection of Human Norovirus via PAMAM Dendrimer/SA-Biotin Mediated Cascade-Amplification.
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 pre-treatment 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 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% (P < 0.01) and 18.09% (P < 0.05) compared with the first two methods, respectively. Three methods were all applied in detecting norovirus in 44 retail oysters, the detection rate of the PAMAM dendrimer/SA-biotin-mediated method was 25.0%, which was higher than those of traditional IME (15.90%) and SA-biotin-amplified IME (18.80%) by 9.1 and 6.2%, respectively. In conclusion, the novel method can be applied for the rapid detection of norovirus in oysters, which can help reduce the cost and time of detection and improve detection rates.
['Gao|Junshan|J|', 'Zhang|Le|L|', 'Xue|Liang|L|', 'Cai|Weicheng|W|', 'Qin|Zhiwei|Z|', 'Yang|Jiale|J|', 'Liang|Yanhui|Y|', 'Wang|Linping|L|', 'Chen|Moutong|M|', 'Ye|Qinghua|Q|', 'Li|Ying|Y|', 'Wang|Juan|J|', 'Wu|Shi|S|', 'Wu|Qingping|Q|', 'Zhang|Jumei|J|']
immunomagnetic enrichment oysters PAMAM-mediated biotin amplification noroviruses