APPLICATION OF MULTIPLEX PCR FOR ANALYSIS OF POLYCOMPONENT PROBIOTIC PREPARATIONS


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Abstract

Aim. Development of multiplex PCR for indication and differential analysis of CFU in a probiotic preparation containing 3 various species of bifidobacteria (Bifidobacterium bifidum, B. longum, B. adolescentis) and 2 species of lactobacilli (Lactobacillus acidophilus, L. plantarum). Materials and methods. Multiplex PCR with primers to 16S RNA genes of 5 species of bacteria was applied. Samples of bacterial colonies were removed from solid nutrient medium and transferred into test-tube for multiplex PCR execution. Bacterial lysates were used as target-matrix for amplification and determination of presence of the required amplicons. Results. Multiplex PCR for detection of CFU of each of the bifidobacteria and lactobacilli strains in samples of 5-component probiotic with accuracy up to species was developed. A reagent kit was fine-tuned and high reproducibility and specificity of the multiplex PCR method with the accuracy of 90-95% of cases was shown. Conclusion. The multiplex PCR method developed is recommended to be applied for CFU control of each bacterial component in a complex probiotic preparation.

About the authors

I. I Vainshtok

Partner Closed Joint-Stock Company, Moscow, Russia

I. Yu Schit

State Scientific Centre of Applied Microbiology and Biotechnology, Obolensk, Russia

V. M Bondarenko

Gamaleya Research Institute of Epidemiology and Microbiology, Moscow, Russia

I. B Brodsky

Partner Closed Joint-Stock Company, Moscow, Russia

References

  1. Бондаренко В.М. Поликомпонентные пробиотики: механизм действия и терапевтический эффект при дисбиозах кишечника. Фарматека. 2005, 20: 46-54.
  2. Бондаренко В.М. Обоснование и тактика назначения в медицинской практике различных форм пробиотических препаратов. Фарматека. 2012, 11: 89-99.
  3. Бондаренко В.М., Мацулевич Т.В. Дисбактериоз кишечника как клинико-лабораторный синдром: современное состояние проблемы. ГЭОТАР-медиа. М., 2007.
  4. Falagas M.E., Betsi G.I., Athanasiou S. Probiotics for prevention of recurrent vulvovaginal candidiasis: A review. J. Antimicrob. Chemother. 2006, 58: 266-272.
  5. FAO/WHO. Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Report of a joint FAO/WHO expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Basel, Switzerland, WHO, 2001.
  6. Kwon H.S., Yang E.H., Yeon S.W, Kang B.H., Kim T.Y. Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA. FEMS Microbiol. Let. 2004, 239: 267-275.
  7. Kwon H.S., Yang E.H., Yeon S.W, Kang B.H., Kim T.Y Rapid identification of potentially probiotic Bifidobacterium species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA. FEMS Microbiol. Let. 2005, 250: 55-62.
  8. Logan A.C., Katzman M. Major depressive disorder: probiotics may be an adjuvant therapy. Med. Hypotheses. 2005, 64 (3): 533-538.
  9. Matsuki T., Watanabe K., Tanaka R., Fukuda M., Oyaizu H. Distribution of bifidobacterial species in human intestinal microflora examined with 16S rRNA-gene-targeted species-specific primers. Appl. Environ. Microbiol. 1999, 65: 4506-4512.
  10. Matsuki T., Watanabe K., Tanaka R. Genus- and species-specific PCR primers for the detection and identification of bifidobacteria. Curr. Issues Intest. Microbiol. 2003, 4: 61-69.
  11. Mitsuoka T. Bifidobacteria and their role in human health. J. Indust. Microbiol. 1990, 6: 263268.
  12. Preidis G.A., Versalovic J. Targeting the human microbiome with antibiotics, probiotics, and prebiotics. Gastroenterology enters the metagenomics era. Gastroenterology. 2009, 136: 20152031.
  13. Reiff C., Kelly D. Inflammatory bowel disease, gut bacteria and probiotic therapy. Int. J. Med. Microbiol. 2010, 300: 25-33.
  14. Song Y.L., Kato N., Liu C.X., Matsumiya Y., Kato H., Watanabe K. Rapid identification of 11 human intestinal Lactobacillus species by multiplex PCR assays using group- and species-specific primers derived from the 16S-23S rRNA intergenic spacer region and its flanking 23S rRNA. FEMS Microbiol.Let. 2000, 187: 167-173.
  15. Walter J., Tannock G. W., Tilsala-Timisjarvi A., Rodtong S., Loach D., Munro M., Alatossava K. T. Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species-specific PCR primers. Appl. Environ. Microbiol. 2000, 66 (1): 297303.

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Copyright (c) 2013 Vainshtok I.I., Schit I.Y., Bondarenko V.M., Brodsky I.B.

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