Detection and quantification of Bacillus cereus and its spores in raw milk by qPCR,and distinguish Bacillus cereus from other bacteria of the genussBacillus  

作　　者：Lijiao Liang Ping Wang Tianming Qu Xiaomei Zhao Yiqiang Ge Ying Chen 梁丽姣;王娉;曲天铭;赵晓美;葛毅强;陈颖(Chinese Academy of Inspection and Quarantine,Beijing,China;College of Food Science and Engineering,Nanjing University of Finance and Economics,Nanjing,China;College of Food Science and Nutritional Engineering,China Agricultural University,Beijing,China;China Rural Technology Development Center,Beijing,China)

机构地区：[1]Chinese Academy of Inspection and Quarantine,Beijing,China [2]College of Food Science and Engineering,Nanjing University of Finance and Economics,Nanjing,China [3]College of Food Science and Nutritional Engineering,China Agricultural University,Beijing,China [4]China Rural Technology Development Center,Beijing,China

出　　处：《Food Quality and Safety》2022年第3期386-395,共10页食品品质与安全研究（英文版）

基　　金：supported by the National Key Research and Development Program of China (2018YFC1604201 and 2018YFC1603800).

摘　　要：Introduction:Raw milk is the basic raw material of dairy products.Bacillus cereus(B.cereus)is a typical conditional pathogenic bacteria and cold-phagocytic spoilage bacteria in raw milk.Materials and Methods:In this study,a quantitative polymerase chain reaction(qPCR)method for detecting B.cereus in raw milk was established.The specificity of the method was verified by using other Bacillus bacteria and pathogenic bacteria;the sensitivity of the method was evaluated by preparing recombinant plasmids and simulated contaminated samples;and the applicability of the method was verified using pure spore DNA.The actual sample detection was completed by using the established qPCR method.Results:The qPCR established in this study can specifically detect B.cereus in raw milk.The limit of detection of the method was as low as 200 CFU/mL,the limit of quantification ranged from 2×10^(2)to 2×10^(8)CFU/mL,and the amplification efficiency of qPCR was 96.6%.Conclusions:The method established in this study can distinguish B.cereus from other Bacillus bacteria,and spore DNA can be used as the detection object.This method has the advantages of strong specificity,high sensitivity,wide application range,and short detection time,which isexpectedtobeapplied in thedairy industry.

关 键 词：Raw milk quality control Bacillus cereus detection SYBR Green real-time qPCR spore DNA detection 

分 类 号：Q93[生物学—微生物学]