Journal
FRONTIERS IN MICROBIOLOGY
Volume 12, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2021.673872
Keywords
noroviruses; oysters; PAMAM-mediated; Immunomagnetic enrichment; biotin amplification
Categories
Funding
- National Key Research and Development Program of China [2018YFC1602500]
- National Natural Science Foundation of China [31872912]
- Natural Science Foundations of Guangdong Province for Distinguished Young Scholars [2019B151502065]
- Key Research and Development Program of Guangdong Province [2019B020209001]
- GDAS' Project of Science and Technology Development [2020GDASYL-20200104008]
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A novel immunomagnetic enrichment method using PAMAM dendrimer/SA-biotin-mediated cascade amplification was established for the detection of norovirus in oysters, showing significantly improved capture efficiency compared to traditional methods. This new method can help reduce costs, save time, and enhance detection rates.
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 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.
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