Evaluating the qualities of probiotic products by a DGGE-based procedure

Authors

  • Tran My Hanh VNU University of Science, Faculty of Biology
  • Cao Thi Dung VNU University of Science, Faculty of Biology
  • Tran Thi Thanh Huyen
  • Pham The Hai

DOI:

https://doi.org/10.15625/1811-4989/17/3/13366

Keywords:

16S rDNA, Denaturing Gradient Gel Electrophoresis (DGGE), probiotic, isolation of microorganisms, DNA extraction

Abstract

The quality of the probiotics is determined by the microbiological composition and can thus be verified by analyzing this characteristic. The use of the Denaturing Gradient Gel Electrophoresis (DGGE) allows the detection of each microorganism through the corresponding band on the gel, which can be considered as the "bar code" of that microorganism. This "bar code” principle demonstrates the potential for the application of DGGE technology to test microbial probiotics with or without a bacterium as registered to assess whether they are qualified. As such, in this study, we experiment the application of DGGE method to test 10 commercial probiotic products, with a focus on products consisting of only bacteria which dominate the market. First, we performed DNA genome extraction and amplification of 550 bp DNA of the 16S rDNA by PCR. Then, the  amplicon was analysed on polyacrylamide gels containing a 30% to 60% linear denaturing gradient of urea and formamide. Microbial composition of 7 of the 10 products showed a strong correlation between DGGE results with product labeling as well as isolation of micoorganism. This study thus demonstrates the potential application of DGGE technology in evaluating the qualities of probiotic products.

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Author Biographies

Tran My Hanh, VNU University of Science, Faculty of Biology

Department of Microbiology

Cao Thi Dung, VNU University of Science, Faculty of Biology

Department of Microbiology

References

Amann RI, Ludwig W, Schleifer KH (1995) Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59(1): 143-169.

Araya M, Morelli L, Reid G, Sanders ME, Stanton C (2002) Joint FAO/WHO Working Group Report on Guidelines for the evaluation of probiotics in food. Food Nutr Sci 5(18).

Azizpour K, Bahrambeygi S, Mahmoodpour S (2009) History and basic of probiotics. Research Journal of Biological Sciences 4(4): 409-426.

Bermudez-Brito M et al. (2012) Probiotic Mechanisms of Action. Ann Nutr Metab 61(2): 160-174.

Chakravorty Soumitesh et al. (2007) A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria. J Microbiol Methods 69(2): 330-339.

Dunbar J, Lawrence OT, Cheryl RK (2000) Assessment of microbial diversity in four southwestern United States soils by 16S rRNA gene terminal restriction fragment analysis. Appl Environ Microbiol 66(7): 2943-2950.

Đỗ Thị Thu Hồng (2017) Nghiên cứu hoàn thiện chế phẩm vi sinh vật ứng dụng cho phân hủy kị khí trong môi trường nước biển. Đại học Khoa học tự nhiên.

Guarner F, Schaafsma GJ (1998) Probiotics. Inl J Food Microbiol 39(3): 237-238.

Hamilton-Miller JM (2004) Probiotics and prebiotics in the elderly. Postgrad Med J 80(946): 447-451.

Hanna S, Fordymacka A, Bardowski J, Gorecki RK, Mrukowicz JZ and Banaszkiewicz A (2004) Microbiological and genetic analysis of probiotic products licensed for medicinal purposes. Med Sci Monit 10: 346-350.

Holben William E et al., (2004) GC Fractionation Enhances Microbial Community Diversity Assessment and Detection of Minority Populations of Bacteria by Denaturing Gradient Gel Electrophoresis. Appl Environ Microbiol 70(4): 2263-2270.

Jacobsen C Nexmann et al., (1999) Screening of probiotic activities of forty-seven strains of Lactobacillus spp. by in vitro techniques and evaluation of the colonization ability of five selected strains in humans. Appl Environ Microbiol 65(11): 4949-4956.

Lopez Isabel et al., (2003) Design and Evaluation of PCR Primers for Analysis of Bacterial Populations in Wine by Denaturing Gradient Gel Electrophoresis. Appl Environ Microbiol 69(11): 6801-6807.

Muyzer G (1999) DGGE/TGGE a method for identifying genes from natural ecosystems. Curr Opin Microbiol 2(3): 317-322.

Muyzer G, E. C. de Waal, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59(3): 695-700.

Nghiêm Ngọc Minh (2005) Sử dụng kỹ thuật điện di trên gel gradient biến tính để nghiên cứu đa dạng vi sinh vật. Tạp chí Công nghệ Sinh học. 2(4): 397-406.

Nguyễn Lân Dũng et al. Khái quát về các phương pháp phân loại vi sinh vật truyền thống. Giáo trình vi sinh vật học. VOER.

Nguyễn Thị Huyền et al. (2014) Khảo sát thành phần vi sinh và các đặc tính probiotic của các sản phẩm men tiêu hóa trên thị trường. Tạp chí Khoa học và Phát triển 12(1).

Parker RB (1974) Probiotics, the other half of the antibiotics story. Anim Nutr Health 29: 4-8.

Rasmussen HB (2012) Restriction fragment length polymorphism analysis of PCR-amplified fragments (PCR-RFLP) and gel electrophoresis-valuable tool for genotyping and genetic fingerprinting. In Gel electrophoresis-principles and Basics. InTech.

Savelkoul PHM et al., (1999) Amplified-fragment length polymorphism analysis: the state of an art. J Clin Microbiol 37(10): 3083-3091.

Schwieger Frank, Christoph C Tebbe (1998) A new approach to utilize PCR–single-strand-conformation polymorphism for 16S rRNA gene-based microbial community analysis. Appl Environ Microbiol 64(12): 4870-4876.

Shah NP (2000) Probiotic bacteria: selective enumeration and survival in dairy foods. J Dairy Sci 83(4): 894-907.

Theunissen J et al,. 2005. Identification of probiotic microorganisms in South African products using PCR-based DGGE analysis. Int J Food Microbiol. 98, 11-21.

Trần Cát Đông (2015) Xu hướng nghiên cứu và ứng dụng chủng lợi khuẩn probiotic trong y học và thực phẩm chức nămg. Sở Khoa học và Công nghệ thành phố Hồ Chí Minh.

Trần Thị Ái Liên (2014) Nghiên cứu đặc điểm và vai trò của Lactobacillus acidophilus trong chế phẩm probiotic. Trường Đại học Sư phạm Thành phố Hồ Chí Minh.

Ward David M et al., (1992) Ribosomal RNA analysis of microorganisms as they occur in nature. Adv Microb Ecol 1992, Springer. 219-286.

Weisburg William G et al., (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173(2): 697-703.

Yeung PS et al., (2002) Species-Specific Identification of Commercial Probiotic Strains. J Dairy Sci 85(5): 1039-1051.

Yaeshima T, Takahashi S, Ishibashi N, and Shimamura S (1996) Identification of bifidobacteria from dairy products and evaluation of microplate hybridization method. Int J Food Microbiol 30: 303–313

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Published

2020-11-28

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