食品科学2017，Vol.38Issue(8)：271-276,6.DOI:10.7506/spkx1002-6630-201708042
EMA-PCR法快速检测啤酒中腐败短乳杆菌
Development of an Ethidium Bromide Monoazide-Polymerase Chain Reaction Assay for Raid Detection of the Beer Spoilage Bacterium Lactobacillus brevis
摘要
Abstract
In this paper,a rapid method using conventional PCR after ethidium bromide monoazide (EMA) pretreatment is described for the detection ofLactobacillus brevis as a beer spoilage bacterium.The PCR amplification was carried out using the hop resistance gene horC as target gene and the genomic DNA from L.brevis as template.The results suggested that addition of EMA to a final concentration lower than 20 μg/mL for pretreatment did not strongly inhibit the PCR amplification of DNA derived from viable L.brevis cells,but it,when added to a final concentration of 1.0 μg/mL,completely inhibited the PCR amplification of DNA derived from 105 CFU/mL dead L.brevis cells.The detection limit (LOD) of EMA-PCR assay was found to be 104 CFU/mL beer sample for the horC gene.Moreover,the horC-specific EMA-PCR assay was applied to detect 13 strains of lactic acid bacteria,representing 100％ specificity with no false positive amplification observed for five beer spoilage lactic acid bacteria and enabling discrimination between the live and dead cells.Overall,the use of horC-specific EMA-PCR allows for a substantial reduction in the rate of false-positive results for potential beer spoilage L.brevis.关键词
叠氮溴乙锭/聚合酶链式反应/啤酒腐败菌/短乳杆菌/酒花耐受基因Key words
ethidium bromide monoazide (EBM)/polymerase chain reaction (PCR)/beer spoilage bacteria/Lactobacillus brevis/horC分类
轻工纺织引用本文复制引用
马艳琳,徐振波,刘君彦,汪东风,邓阳..EMA-PCR法快速检测啤酒中腐败短乳杆菌[J].食品科学,2017,38(8):271-276,6.基金项目
中国博士后科学基金资助项目(2015M582063 （）
2014T70810) （）
南昌大学食品科学与技术国家重点实验室开放基金资助项目(SKLF-KF-201415) （SKLF-KF-201415）
