Droplet Digital PCR Is an Improved Alternative Method for High-Quality Enumeration of Viable Probiotic Strains

Frontiers in Microbiology

Volumn 10, Issue , 2020, Pages

  Hansen, Sarah J Z ^a   Tang, Peipei ^a   Kiefer, Anthony ^a   Galles, Kevin ^a   Wong, Connie ^a   Morovic, Wesley ^a  

^a DUPONT COMPANY   (United States)

Author keywords

B. animalis subsp. lactis HN019; Bifidobacterium animalis subsp. lactis Bl 04; droplet digital PCR; enumeration; flow cytometry; Lactobacillus acidophilus NCFM; plate count; probiotic

Indexed keywords

PROBIOTIC AGENT;

ARTICLE; BIFIDOBACTERIUM ANIMALIS; CELL COUNTING; CELL VIABILITY; COLONY FORMING UNIT; COMPARATIVE STUDY; COPY NUMBER VARIATION; CORRELATION ANALYSIS; DROPLET DIGITAL POLYMERASE CHAIN REACTION; FLOW CYTOMETRY; LACTOBACILLUS ACIDOPHILUS; NONHUMAN; PLATE COUNT;

EID: 85078979248     PISSN: None     EISSN: 1664302X     Source Type: Journal    
DOI: 10.3389/fmicb.2019.03025     Document Type: Article

References (28)

1. Bland J. M., Altman D. G., (1986). Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1 307–310. 10.1016/s0140-6736(86)90837-8 2868172
2. Briczinski E. P., Loquasto J. R., Barrangou R., Dudley E. G., Roberts A. M., Roberts R. F., (2009). Strain-Specific Genotyping of Bifidobacterium animalis subsp. lactis by Using Single-Nucleotide Polymorphisms. Insertions, and Deletions. Appl. Environ. Microbiol. 75 7501–7508. 10.1128/AEM.01430-1439 19801460
3. Codony F., Agustí G., Allué-Guardia A., (2015). Cell membrane integrity and distinguishing between metabolically active and inactive cells as a means of improving viability PCR. Mol. Cell. Probes 29 190–192. 10.1016/j.mcp.2015.03.003 25797787
4. Davis C., (2014). Enumeration of probiotic strains: review of culture-dependent and alternative techniques to quantify viable bacteria. J. Microbiol. Methods 103 9–17. 10.1016/j.mimet.2014.04.012 24814752
5. Fenster K., Freeburg B., Hollard C., Wong C., Rønhave Laursen R., Ouwehand A. C., (2019). The Production and Delivery of Probiotics: a Review of a Practical Approach. Microorganisms 7: pii: E83. 10.3390/microorganisms7030083 30884906
6. ® via chip-based digital pcr. Front. Microbiol. 9:704. 10.3389/fmicb.2018.00704 29696008
7. Heller K. J., (2001). Probiotic bacteria in fermented foods: product characteristics and starter organisms. Am. J. Clin. Nutr 73 374s–379s. 10.1093/ajcn/73.2.374s 11157344
8. Herbel S. R., Lauzat B., von Nickisch-Rosenegk M., Kuhn M., Murugaiyan J., Wieler L. H., et al. (2013). Species-specific quantification of probiotic lactobacilli in yoghurt by quantitative real-time PCR. J. Appl. Microbiol. 115 1402–1410. 10.1111/jam.12341 24024971
9. Hill C. R., Guarner F., Reid G., Gibson G. F. R., Merenstein D. J., Pot B., et al. (2014). Expert consensus document: the International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nature Rev. Gastroenterol. Hepatol. 11 506–514. 10.1038/nrgastro.2014.66 24912386
10. ISO (2006). ISO 20128:2006. ISO. Available at: http://www.iso.org/cms/render/live/en/sites/isoorg/contents/data/standard/03/52/35292.html (accessed June 3, 2019).
11. ISO (2010). ISO 29981:2010. ISO. Available at: http://www.iso.org/cms/render/live/en/sites/isoorg/contents/data/standard/04/57/45765.html (accessed June 3, 2019).
12. Iso (2015). ISO 19344:2015 - Milk and milk products – Starter cultures, probiotics and fermented products – Quantification of lactic acid bacteria by flow cytometry. Available at: https://www.iso.org/standard/64658.html (accessed June 3, 2019).
13. Jackson S. A., Schoeni J. L., Vegge C., Pane M., Stahl B., Bradley M., et al. (2019). Improving End-User Trust in the Quality of Commercial Probiotic Products. Front. Microbiol. 10:739. 10.3389/fmicb.2019.00739 31105649
14. Kramer M., Obermajer N., Bogovič Matijašić B., Rogelj I., Kmetec V., (2009). Quantification of live and dead probiotic bacteria in lyophilised product by real-time PCR and by flow cytometry. Appl. Microbiol. Biotechnol. 84 1137–1147. 10.1007/s00253-009-2068-2067 19529931
15. Luedtke B. E., Bosilevac J. M., (2015). Comparison of methods for the enumeration of enterohemorrhagic Escherichia coli from veal hides and carcasses. Front. Microbiol. 6:1062. 10.3389/fmicb.2015.01062 26483780
16. Lugli G. A., Mangifesta M., Mancabelli L., Milani C., Turroni F., Viappiani A., et al. (2019). Compositional assessment of bacterial communities in probiotic supplements by means of metagenomic techniques. Int. J. Food Microbiol. 294 1–9. 10.1016/j.ijfoodmicro.2019.01.011 30711887
17. Morovic W., Hibberd A. A., Zabel B., Barrangou R., Stahl B., (2016). Genotyping by PCR and High-Throughput Sequencing of Commercial Probiotic Products Reveals Composition Biases. Front. Microbiol. 7:1747. 10.3389/fmicb.2016.01747 27857709
18. Morovic W., Roos P., Zabel B., Hidalgo-Cantabrana C., Kiefer A., Barrangou R., (2018). Transcriptional and Functional Analysis of Bifidobacterium animalis subsp. lactis Exposure to Tetracycline. Appl. Environ. Microbiol. 84 e1999–e1918. 10.1128/AEM.01999-1918 30266728
19. ® Restore (Lactobacillus acidophilus NCFM. Lactobacillus paracasei Lpc-37, Bifidobacterium animalis subsp. lactis Bl-04 and B. lactis Bi-07) for antibiotic resistance, genomic risk factors, and acute toxicity. Food Chem. Toxicol. 110 316–324. 10.1016/j.fct.2017.10.037 29080807
20. Mukaka M., (2012). A guide to appropriate use of Correlation coefficient in medical research. Malawi Med. J. 24 69–71. 23638278
21. ® (Lactobacillus plantarum CECT 7527, 7528 and 7529): antibiotic resistance and 90-day repeated-dose study in rats. Food Chem. Toxicol. 92 117–128. 10.1016/j.fct.2016.03.018 27016492
22. Naghili H., Tajik H., Mardani K., Razavi Rouhani S. M., Ehsani A., Zare P., (2013). Validation of drop plate technique for bacterial enumeration by parametric and nonparametric tests. Vet. Res. Forum 4 179–183. 25653794
23. Ouwehand A. C., Invernici M. M., Furlaneto F. A. C., Messora M. R., (2018). Effectiveness of multi-strain versus single-strain probiotics: current status and recommendations for the future. J. Clin. Gastroenterol. 52 s35–s40. 10.1097/MCG.0000000000001052 29734210
24. Ouwehand A. C., Tölkkö S., Kulmala J., Salminen S., Salminen E., (2000). Adhesion of inactivated probiotic strains to intestinal mucus. Lett. Appl. Microbiol. 31 82–86. 10.1046/j.1472-765x.2000.00773.x 10886621
25. Patro J. N., Ramachandran P., Barnaba T., Mammel M. K., Lewis J. L., Elkins C. A., (2016). Culture-Independent Metagenomic Surveillance of Commercially Available Probiotics with High-Throughput Next-Generation Sequencing. mSphere 1:e00057-16. 10.1128/mSphere.00057-16 27303722
26. Pegg D. E., (1989). Viability assays for preserved cells, tissues, and organs. Cryobiology 26 212–231. 10.1016/0011-2240(89)90016-90013
27. Villarreal M. L. M., Padilha M., Vieira A. D. S., Franco B. D. G., Martinez R. C. R., Saad S. M. I., (2013). Advantageous Direct Quantification of Viable Closely Related Probiotics in Petit-Suisse Cheeses under In Vitro Gastrointestinal Conditions by Propidium Monoazide - qPCR. PLoS ONE 8:e82102. 10.1371/journal.pone.0082102 24358142
28. Wilkinson M. G., (2018). Flow cytometry as a potential method of measuring bacterial viability in probiotic products: a review. Trends Food Sci. Technol. 78 1–10. 10.1016/j.tifs.2018.05.006