Quantification and discovery of PCR inhibitors found in food matrices commonly associated with foodborne viruses  

作　　者：Cassandra Suther Matthew D.Moore 

机构地区：[1]Department of Food Science,University of Massachusetts,Amherst,MA 01003,United States

出　　处：《Food Science and Human Wellness》2019年第4期351-355,共5页食品科学与人类健康（英文）

基　　金：This project was funded by the University of MassachusettsAmherst.

摘　　要：Human norovirus is the leading cause of foodborne illness globally.Detection and quantification of norovirus commonly involves the use of reverse transcriptase quantitative polymerase chain reaction(RT-qPCR);however,the presence of inhibitory compounds in foods limit detection and accurate quantification.Although some studies have been done on PCR inhibitors from foods,many of them are over a decade old and do not investigate inhibition in contemporary one-step RT-qPCR-based detection chemistries.The purpose of this work was to quantify the degree of inhibition that occurs from inhibitory compounds found in produce(pectin)and mollusks(hemocyanin,glycogen)—foods commonly associated with norovirus outbreaks.RT-qPCR reactions containing different amounts of genomic bacteriophage MS2 RNA,a norovirus surrogate,were spiked with different concentrations of pectin(0.0625%–0.25%w/V),glycogen(1.25%–10%),and hemocyanin(0.0625%–0.25%).Past research has implicated glycogen as an inhibitory compound in oysters;however,even high levels of glycogen(10%)had no significant effect(P>0.05)on amplification.Conversely,both pectin and hemocyanin caused complete inhibition at 0.25%,with no significant inhibition observed at 0.0625%(P<0.05).Hemocyanin is abundant in the hemolymph of mollusks and previously untested as a PCR inhibitor.This work demonstrates that pectin and hemocyanin should be considered when testing produce and mollusk samples with PCR-based methods.

关 键 词：PCR inhibition PCR false negatives Norovirus detection HEMOCYANIN PECTIN 

分 类 号：TS207.3[轻工技术与工程—食品科学]