Establishment and preliminary application of an efficient nanomaterial-assisted PCR assay for detection of astrovirus in shellfish aquatic products
Human astrovirus(HAstV)is one of the main pathogens causing global diarrhea.HAstV mainly presents in shellfishes.Shellfishes are filter-feeding animals and their digestive tissues accumulated and enriched viruses in water.It can make them as the carriers of important transmission.Consumers were mainly infected with HAstV through processes of processing and cooking shellfishes.A tiny amount of virus can cause infection in young children and patients with acquired immune dysfunction.The patients were accompanied by clinical symptoms,such as diarrhea and vomiting.In order to timely detect HAstV in shellfishes and ensure safety of consumers,we established a simple,sensitive and specific method.The nanometer materials were integrated into PCR.We designed specific primers for the conserved HAstV ORF2 gene.By optimizing the content of AuNPs,the concentration of primers and the annealing temperature,we established an Nanomaterial-assisted PCR(NanoPCR)method to detect HAstV.This study indicated that the HAstV NanoPCR assay has characteristics of high sensitivity and specificity.The minimum of viral detection line was 2.0 × 100 copies/μL,the sensitivity of NanoPCR was 1 000 times higher than traditional PCR.There was no cross reactions with other viruses in shellfishes.Using NanoPCR method to detect HAstV,the detection rate was 7.5％in shellfishes,while the detection rate of traditional PCR was 5.35％.Research has shown that the HAstV NanoPCR method is suitable for screening HAstV in shellfishes.It can be timely applied to detect the prevalence of HAstV in aquatic products and ensure to provide technical support for food safety of consumers.
万方数据
维普
