Quantitative detection of gii-type norovirus in fruits and vegetables by one-step microdroplet digital PCR

Abstract

The invention discloses a one-step droplet digital PCR method for quantitatively detecting the GII type norovirus in fruits and vegetables. According to the method, RT-ddPCR is utilized, a primer and a probe for RT-qPCR detection of the GII type norovirus are combined for use, reaction is carried out, and then the high-sensitivity rapid detection for the GII type norovirus is realized. The method is high in sensitivity and can achieve about 2copies / [mu] L, but the sensitivity of the conventional RT-qPCR is usually 10 copies / [mu] L; the amplification efficiency is high, the amplification efficiency is 95.4%, R2 is equal to 0.9973. Under low copy number, compared with RT-qPCR, the method provided by the invention can more effectively avoid the influences of inhibitors in fruits and vegetables, and the false negative generation is reduced. Therefore, the detection method provided by the invention is sensitive, accurate and visual, a novel detection method is provided for the government departments including the agriculture department and the inspection and quarantine bureau, and related detection mechanisms and enterprises, and guiding significance is also achieved.

Description

technical field

[0001] The invention relates to the technical field of food quality and safety detection, in particular to a one-step microdrop digital PCR quantitative detection method for GII type norovirus in fruits and vegetables. Background technique

[0002] Foodborne viral illness is a social problem that is receiving attention. Worldwide, more than 50% of acute gastroenteritis events are caused by norovirus, the main genotype of which is GII. Norovirus was first detected by Kapikan et al. in 1972. It has the characteristics of low infectious dose and can tolerate a wide range of temperature changes. It is an important cause of non-bacterial acute gastroenteritis. Its main route of transmission is "fecal-oral", which can be transmitted directly from person to person, or through contact with contaminated water and food. Frequently contaminated foods include vegetables, soft fruits, and shellfish. Wait. How to detect and quantify norovirus in food is a hot issue. C...

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