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Use of tuf gene-based primers for the PCR detection of probiotic Bifidobacterium species and enumeration of bifidobacteria in fermented milk by cultural and quantitative real-time PCR methods.
J Food Sci. 2010 Oct; 75(8):M521-7.JF

Abstract

Due to the increasing use of bifidobacteria in probiotic products, it is essential to establish a rapid method for the qualitative and quantitative assay of the bifidobacteria in commercial products. In this study, partial sequences of the tuf gene for 18 Bifidobacterium strains belonging to 14 species were determined. Alignment of these sequences showed that the similarities among these Bifidobacterium species were 82.24% to 99.72%. Based on these tuf gene sequences, 6 primer sets were designed for the polymerase chain reaction (PCR) assay of B. animalis subsp. animalis, B. animalis subsp. lactis, B. bifidum, B. breve, B. longum subsp. infantis, B. longum subsp. longum, and the genus of Bifidobacterium, respectively. These Bifidobacterium species are common probiotic species present in dairy and probiotic products. When each target Bifidobacterium spp. was assayed with the designed primers, PCR product with expected size was generated. In addition, for each target species, more than 70 bacterial strains other than the target species, including strains of other Bifidobacterium species, strains of Lactobacillus spp., Enterococcus spp., and other bacterial species, all generated negative results. PCR assay with primers specific to B. animalis subsp. lactis and B. longum subsp. longum confirmed the presence of these Bifidobacterium species in commercial yogurt products. In addition, for each product, enumeration of the bifidobacteria cells by culture method with BIM-25 agar and the quantitative real-time PCR showed similar cell counts. Such results indicated that within 15-d storage (4 °C) after manufacture, all the bifidobacteria cells originally present in yogurt products were viable and culturable during the storage.

Authors+Show Affiliations

Dept. of Food Science and Biotechnology, Natl. Chung Hsing Univ., No. 250, Kuo Kuang Rd. Taichung City, Taiwan 402, Republic of China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21535508

Citation

Sheu, Sen-Je, et al. "Use of Tuf Gene-based Primers for the PCR Detection of Probiotic Bifidobacterium Species and Enumeration of Bifidobacteria in Fermented Milk By Cultural and Quantitative Real-time PCR Methods." Journal of Food Science, vol. 75, no. 8, 2010, pp. M521-7.
Sheu SJ, Hwang WZ, Chiang YC, et al. Use of tuf gene-based primers for the PCR detection of probiotic Bifidobacterium species and enumeration of bifidobacteria in fermented milk by cultural and quantitative real-time PCR methods. J Food Sci. 2010;75(8):M521-7.
Sheu, S. J., Hwang, W. Z., Chiang, Y. C., Lin, W. H., Chen, H. C., & Tsen, H. Y. (2010). Use of tuf gene-based primers for the PCR detection of probiotic Bifidobacterium species and enumeration of bifidobacteria in fermented milk by cultural and quantitative real-time PCR methods. Journal of Food Science, 75(8), M521-7. https://doi.org/10.1111/j.1750-3841.2010.01816.x
Sheu SJ, et al. Use of Tuf Gene-based Primers for the PCR Detection of Probiotic Bifidobacterium Species and Enumeration of Bifidobacteria in Fermented Milk By Cultural and Quantitative Real-time PCR Methods. J Food Sci. 2010;75(8):M521-7. PubMed PMID: 21535508.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Use of tuf gene-based primers for the PCR detection of probiotic Bifidobacterium species and enumeration of bifidobacteria in fermented milk by cultural and quantitative real-time PCR methods. AU - Sheu,Sen-Je, AU - Hwang,Wen-Zhe, AU - Chiang,Yu-Cheng, AU - Lin,Wen-Hsin, AU - Chen,Hsin-Chih, AU - Tsen,Hau-Yang, PY - 2011/5/4/entrez PY - 2011/5/4/pubmed PY - 2011/8/24/medline SP - M521 EP - 7 JF - Journal of food science JO - J Food Sci VL - 75 IS - 8 N2 - Due to the increasing use of bifidobacteria in probiotic products, it is essential to establish a rapid method for the qualitative and quantitative assay of the bifidobacteria in commercial products. In this study, partial sequences of the tuf gene for 18 Bifidobacterium strains belonging to 14 species were determined. Alignment of these sequences showed that the similarities among these Bifidobacterium species were 82.24% to 99.72%. Based on these tuf gene sequences, 6 primer sets were designed for the polymerase chain reaction (PCR) assay of B. animalis subsp. animalis, B. animalis subsp. lactis, B. bifidum, B. breve, B. longum subsp. infantis, B. longum subsp. longum, and the genus of Bifidobacterium, respectively. These Bifidobacterium species are common probiotic species present in dairy and probiotic products. When each target Bifidobacterium spp. was assayed with the designed primers, PCR product with expected size was generated. In addition, for each target species, more than 70 bacterial strains other than the target species, including strains of other Bifidobacterium species, strains of Lactobacillus spp., Enterococcus spp., and other bacterial species, all generated negative results. PCR assay with primers specific to B. animalis subsp. lactis and B. longum subsp. longum confirmed the presence of these Bifidobacterium species in commercial yogurt products. In addition, for each product, enumeration of the bifidobacteria cells by culture method with BIM-25 agar and the quantitative real-time PCR showed similar cell counts. Such results indicated that within 15-d storage (4 °C) after manufacture, all the bifidobacteria cells originally present in yogurt products were viable and culturable during the storage. SN - 1750-3841 UR - https://www.unboundmedicine.com/medline/citation/21535508/Use_of_tuf_gene_based_primers_for_the_PCR_detection_of_probiotic_Bifidobacterium_species_and_enumeration_of_bifidobacteria_in_fermented_milk_by_cultural_and_quantitative_real_time_PCR_methods_ DB - PRIME DP - Unbound Medicine ER -