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

作     者：Cassandra Suther Matthew D.Moore 

作者机构：Department of Food ScienceUniversity of MassachusettsAmherstMA 01003United States 

出 版 物：《Food Science and Human Wellness》 (食品科学与人类健康（英文）)

年 卷 期：2019年第8卷第4期

页      面：351-355页

核心收录：

学科分类：0832[工学-食品科学与工程（可授工学、农学学位）] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 09[农学] 0903[农学-农业资源与环境] 

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

主　　题：PCR inhibition PCR false negatives Norovirus detection Hemocyanin Pectin 

摘      要：Human norovirus is the leading cause of foodborne illness *** 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 *** 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 *** 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 ***-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(P0.05)on ***,both pectin and hemocyanin caused complete inhibition at 0.25%,with no significant inhibition observed at 0.0625%(P0.05).Hemocyanin is abundant in the hemolymph of mollusks and previously untested as a PCR *** work demonstrates that pectin and hemocyanin should be considered when testing produce and mollusk samples with PCR-based methods.