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

• Published in: Frontiers in microbiology Vol. 12; p. 673872
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: Frontiers Media S.A 20-07-2021
• Subjects: biotin amplification; immunomagnetic enrichment; Microbiology; noroviruses; oysters; PAMAM-mediated
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2021.673872

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.