食品与发酵工业2018，Vol.44Issue(2)：194-201,8.DOI:10.13995/j.cnki.11-1802/ts.015327
实时荧光定量PCR法定量微生物的条件及在鱼露和虾油微生物检测中的应用研究
Determination of the conditions for the counting of microbes by real-time fluorescent quantitative PCR and research on the detection of microbial populations in fish sauce and shrimp sauce
摘要
Abstract
In this daper,the conditions of determining baterial and fangal populations by real-time fluorescert quantitative PCR (FQ-PCR) method were investigated.By comparing the amplification effects of several sets of primers,the bacterial 16S rDNA universal primer 63F-335R and the fungal 18S rDNA universal primer 0817F-1196R were respectively selected as the primers for FQ-PCR detection of bacterial and fungal populations.And the annealing temperatures were set to be 60 ℃.The selected FQ-PCR primers and the annealing temperatures were validated using many strains of bacteria and fungi.The results showed that for any single or mixed strains,the amplification efficiencies were within 90％-120％ and the determination coefficients of the standard curves were greater than 0.98,which satisfied the requirements of quantification.The bacterial and fungal populations in fish sauce and shrimp sauce weredetected by FQ-PCR under above conditions.The results revealed that during fermentation of fish sauce,the counts ofbacterial population varied within the range of 3.16 × 104-3.16 × 105 cells/ml,which were far higher than those ofthe fungal population.Besides,the counts of bacterial and fungal populations in fish sauce and shrimp sauce deter-mined by the FQ-PCR method were 1-2 orders of magnitude higher than those determined by plate count method.关键词
荧光定量PCR/细菌计数/真菌计数/鱼露/虾油Key words
fluorescent quantitative PCR/counting of bacteria/counting of fungi/fish sauce/shrimp sauce引用本文复制引用
王香君,李梦茹,段杉..实时荧光定量PCR法定量微生物的条件及在鱼露和虾油微生物检测中的应用研究[J].食品与发酵工业,2018,44(2):194-201,8.基金项目
广东省省部产学研合作专项资金项目(2013B090600111) （2013B090600111）
粤港关键领域重点突破项目(2010A020104004) （2010A020104004）
四川省创新能力提升工程项目(No.2016TSCY-006) （No.2016TSCY-006）
