实时荧光定量PCR技术监测腌制麻竹笋中乳酸乳球菌动态变化  

Detection of Lactococcus lactis in Pickled Ma Bamboo Shoots by Quantitative Real-time PCR

作　　者：夏雪娟[1] 郑炯[1,2] 叶秀娟[1] 吴金松[1] 阚建全[1,2] 

机构地区：[1]西南大学食品科学学院,重庆400715 [2]重庆市特色食品工程技术研究中心,重庆400715

出　　处：《食品科学》2016年第4期88-92,共5页Food Science

基　　金：中央高校基本科研业务费专项(XDJK2013C131);重庆市特色食品工程技术研究中心能力提升项目(cstc2014pt-gc8001)

摘　　要：采用实时荧光定量聚合酶链式反应（quantitative real-time polymerase chain reaction,q RT-PCR）技术定量监测6 g/100 m L盐质量浓度腌制麻竹笋中乳酸乳球菌（Lactococcus lactis）的动态变化。经乳酸乳球菌标准菌株基因组DNA提取、标准阳性质粒制备、标准曲线绘制、各时期竹笋腌制发酵液中细菌基因组DNA提取和乳酸乳球菌qRTPCR特异性扩增,对腌制液中乳酸乳球菌进行定量检测。结果表明,在腌制过程中（0-63 d）,随着腌制时间的延长,乳酸乳球菌含量逐渐升高,在腌制14 d时达到最大值（4.63×10^8 copies/μL）,与0 d（2.41×10^2 copies/μL）相比增加了6个数量级,而后浓度缓慢降低,在腌制63 d时浓度为5.02×10^6 copies/μL。qRT-PCR技术为定量监测腌制麻竹笋中微生物的动态变化提供了一条可靠、快速的有效途径。The dynamic changes of Lactococcus lactis in pickled Ma bamboo shoots with 6 g/100 m L salt concentration were studied by quanti tative real-time polymerase chain reaction（q RT-PCR）. After the DNA extraction from standard Lactococcus lactis,preparation of positive plasmids,protraction of standard curve,DNA extraction from samples at different fermentation stages and q RT-PCR,and quantitative detection were performed. The results showed that during fermentation（0–63 days）,the concentration of Lactococcus lactis increased gradually from 2.41×10^2 copies/μL（day 0） to the maximum value of 4.63×10^8 copies/μL（day 14）,an increase by six orders of magnitude,followed by a slow decrease to 5.02×10^6 copies/μL at day 63. In conclusion,q RT-PCR technology provides a reliable,fast and effective way for the quantitative analysis of bacteria in pickled Ma bamboo shoots.

关 键 词：大叶麻竹笋 腌制 实时荧光定量PCR 乳酸乳球菌 动态变化 

分 类 号：TS201.3[轻工技术与工程—食品科学]