Updating and evaluation of nested RT-PCR for specific detection of genogroup Ⅱ norovirus in oysters

Abstract:

New primers (NG2OF/NG2OR) were designed based on analysis of all GⅡ norovirus sequences available online to solve the problem of non-specific amplification and false negative in detecting GⅡ norovirus in oysters by performing the present nested RT-PCR method. The primers are evaluated for their coverage, specificity, sensitivity and stability. The results show that: new primers were specifically targeted for GⅡ norovirus and not affected by the original background of the oyster samples; the assay successfully detects GⅡ norovirus from norovirus contaminated oysters with the sensitivity of 26.4 noroviruses; the assay has a 28.3% positive rate as 92 oyster samples are detected, which is 7.5% higher than the typical method. Thus, the new nested PCR method, with a higher sensitivity, is more efficient in the detection of oyster samples. It provided a new and reliable assay for the detection of GⅡ norovirus in oysters.