食品与发酵工业 >

2020 , Vol. 46 >Issue 15: 238 - 244

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.023849

分析与检测

采用实时荧光定量PCR法同时定性定量检测发酵乳中多种益生菌

  • 覃璐琪 ,
  • 梁晓琳 ,
  • 潘海博 ,
  • 莫明规 ,
  • 饶川艳 ,
  • 聂梦琳 ,
  • 梅丽华 ,
  • 李全阳
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  • 1(广西大学 轻工与食品工程学院,广西 南宁,500004)
    2(柳州市康小乐牛奶有限公司,广西 柳州,545007)
硕士研究生(李全阳教授为通讯作者,E-mail：liquanyang@gxu.edu.cn)

收稿日期: 2020-03-04

  修回日期: 2020-04-16

  网络出版日期: 2020-08-15

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Simultaneously qualitative and quantitative detections of probiotics in fermented milk using qPCR

  • QIN Luqi ,
  • LIANG Xiaolin ,
  • PAN Haibo ,
  • MO Minggui ,
  • RAO Chuanyan ,
  • NIE MengLin ,
  • MEI Lihua ,
  • Li Quanyang
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  • 1(College of Light Industry and Food Engineering, Guangxi University,Nanning 500004,China)
    2(Liuzhou Kangxiaole Dairy Co. Ltd,Liuzhou 545007,China)

Received date: 2020-03-04

  Revised date: 2020-04-16

  Online published: 2020-08-15

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摘要

为快速、准确地对益生菌发酵乳中的干酪乳杆菌及罗伊氏乳杆菌进行定性、定量检测,通过设计干酪乳杆菌及罗伊氏乳杆菌种属特异性引物,建立了双重实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测方法。结果表明,所建标准曲线方程线性关系良好,相关系数R2均达到0.99以上。对市售产品随机抽样检测发现：样品1、样品2未添加干酪乳杆菌及罗伊氏乳杆菌。样品3中检出干酪乳杆菌(1.39±0.25)×109 copies/mL,未检出罗伊氏乳杆菌。自制益生菌发酵乳检出干酪乳杆菌(3.7±0.35)×109 copies/mL、罗伊氏乳杆菌(4.8±0.26)×109 copies/mL,检测结果均与所知的益生菌种类及数量信息相符。结果表明,上述建立的实时荧光定量PCR方法能够同时对益生菌发酵乳中的干酪乳杆菌及罗伊氏乳杆菌进行快速、准确地定性、定量检测,克服了平板培养法和流式细胞术的局限,具有一定的优势和应用前景。

关键词： 干酪乳杆菌; 罗伊氏乳杆菌; 荧光定量PCR; 定性检测; 定量检测

本文引用格式

覃璐琪 , 梁晓琳 , 潘海博 , 莫明规 , 饶川艳 , 聂梦琳 , 梅丽华 , 李全阳 . 采用实时荧光定量PCR法同时定性定量检测发酵乳中多种益生菌[J]. 食品与发酵工业, 2020 , 46(15) : 238 -244 . DOI: 10.13995/j.cnki.11-1802/ts.023849

Abstract

In order to rapidly and accurately detect the quantity and the quality of Lactobacillus casei and Lactobacillus reuteri in probiotic fermented milk,a dual real-time fluorescence quantitative polymerase chain reaction (PCR) detection method was established by selecting and optimizing specific amplification primers of L. casei and L. reuteri. The results showed that the established standard curve equation had a good linear relationship,and the correlation coefficient R2 was above 0.99. Random sampling of commercial products showed that sample 1 and 2 did not contain L. casei or L. reuteri. In DIY probiotics fermented milk (3.7±0.35)×109 copies/mL of L. casei and (4.8±0.26)×109 copies/mL of L. reuteri was detected,while in sample 3 (1.39±0.25)×109 copies/mL of L. casei and no L. reuteri was detected. The detection results were consistent with the known probiotic species and quantity information. The above results showed that the real-time fluorescence quantitative PCR method established in this study could detect the quantity and quality of L. casei and L. reuteri in probiotic fermented milk quickly and accurately.

Key words： Lactobacillus casei; Lactobacillus reuteri; fluorescence quantitative PCR; qualitative detection; quantitative detection

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