摘要
通过对Lactobacillus paracasei种内的20株菌的全基因组序列进行了单核苷酸多态性(single nucleotide polymorphism,SNP)位点分析,并结合糖代谢途径预测,序列比对,得到了分辨力可达菌株水平且遗传相对稳定的N1115菌株特异性位点。针对特异性位点所设计的N1115菌株特异性荧光定量PCR扩增引物及探针组,在51株常见益生菌株间具有特异性,所选特异性位点传代8代比较稳定,针对菌粉所建DNA提取方法的定量检出限为10^(3)~10^(9)CFU/mL,其间的线性拟合度较好(R2>0.98)。采用已知浓度标准菌粉做标准曲线可以对未知菌含量样品进行准确定量。
Based on the SNPs analysis of 20 strains in Lactobacillus paracasei,combined with the prediction of sugar metabolism pathway and sequence alignment,the specific site of N1115 strain with high resolution and relatively stable heredity was obtained.The designed primers and probes were specific among 51 common probiotics,and the selected specific sites were stable in 8 generation.The detection limit of DNA extraction method for bacterial powder was 10^(3)CFU/mL to 10^(9)CFU/mL,and the linear fitting between them was good(R^(2)>0.98).Using the known concentration standard bacterial powder as the standard curve,the unknown bacterial content samples can be accurately quantified.
作者
张奕南
王越
张栋
张娜娜
朱宏
刘洋
王世杰
ZHANG Yinan;WANG Yue;ZHANG Dong;ZHANG Nana;ZHU Hong;LIU yang;WANG Shijie(Shanghai Institute of Quality Inspection and Technical Research,Shanghai 200233,China;The Shijiazhang Junlebao Dairy Co.Ltd.,Shijiazhuang 050221,China)
出处
《中国乳品工业》
CAS
北大核心
2021年第4期45-53,64,共10页
China Dairy Industry
基金
十三五国家重点研发计划(2016YFD0400605)
上海市闵行区产学研项目(2019MHC004)
上海市食品质量安全检测与评价专业技术服务平台(18DZ2292400)。
关键词
副干酪乳杆菌SNP位点
菌株定量检测
益生菌
Lactobacillus paracasei SNPs
quantitative detection of strains
probiotics
