Rapid assay to assess colonization patterns following in-vivo probiotic ingestion

BMC Research Notes

Sepehr N Tabrizi⁴  Marie Pirotta¹  Susan E Jacobs²  Suzanne M Garland⁴  Jacinta M Tobin³ 

[1] Department of General Practice, University of Melbourne, Parkville, VIC 3052, Australia;Neonatal Services, The Royal Women’s Hospital, Parkville, VIC, Australia;Northwest Academic Centre University of Melbourne, Sunshine Hospital, St Albans, VIC 3021, Australia;Murdoch Childrens Research Institute, Parkville, VIC 3052, Australia

关键词: Micro biome;    Preterm infant;    qPCR;    Probiotics;   

Others  :  1142296
DOI  :  10.1186/1756-0500-6-252

 received in 2012-11-06, accepted in 2013-07-03,  发布年份 2013

【 摘 要 】

Background

Colonization of the intestine with some microorganisms has been shown to have beneficial health effects. The association of bacteria with its human host starts soon after birth; however in infants born prematurely establishment of normal intestinal flora is interrupted with colonization with potential pathogenic organisms Probiotic supplementation may therefore be beneficial to the health of preterm infants. As most probiotic organisms are difficult to culture, confirmation of their colonization after supplementation is difficult. In this study, rapid qPCR assays for detection of presence of probiotic species in the intestine by faecal sampling is described in both preterm infant and adult participants.

Findings

Probiotic colonization was determined using qPCR directed at amplification of organisms present in the ingested probiotic Streptococcus thermophilus, Bifidobacterium animalis subsp. lactis and B. longum subsp. infantis. Overall, differential detection of probiotic strains in faeces were found between adult and preterm infants, with 50% of infants continuing to shed at least two probiotic strains three weeks after probiotic ingestion had ceased.

Conclusions

This study demonstrated rapid assessment of the preterm infant gut for colonization with probiotic strains using real-time PCR. This method would be of great importance in studies of probiotics in prevention of diseases and adverse clinical outcomes.

【 授权许可】

   
2013 Tobin et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA: Diversity of the human intestinal microbial flora. Science 2005, 308(5728):1635-1638.
• [2]Hooper LV, Bry L, Falk PG, Gordon JI: Host-microbial symbiosis in the mammalian intestine: exploring an internal ecosystem. Bioessays 1998, 20(4):336-343.
• [3]Mazmanian SK, Liu CH, Tzianabos AO, Kasper DL: An immunomodulatory molecule of symbiotic bacteria directs maturation of the host immune system. Cell 2005, 122(1):107-118.
• [4]Huurre A, Kalliomaki M, Rautava S, Rinne M, Salminen S, Isolauri E: Mode of delivery - effects on gut micro biota and humoral immunity. Neonatology 2008, 93(4):236-240.
• [5]Prescott SL, Bjorksten B: Probiotics for the prevention or treatment of allergic diseases. J Allergy Clin Immunol 2007, 120(2):255-262.
• [6]Penders J, Thijs C, Vink C, Stelma FF, Snijders B, Kummeling I, van den Brandt PA, Stobberingh EE: Factors influencing the composition of the intestinal micro biota in early infancy. Pediatrics 2006, 118(2):511-521.
• [7]Schwiertz A, Gruhl B, Lobnitz M, Michel P, Radke M, Blaut M: Development of the intestinal bacterial composition in hospitalized preterm infants in comparison with breast-fed, full-term infants. Pediatr Res 2003, 54(3):393-399.
• [8]Léké A, Romond MB, Mullié C: Insights in the Human Bifidobacterial Flora Through Culture-Dependent and Independent Techniques. In Communicating Current Research and Educational Topics and Trends in Applied Microbiology, Volume 2. Edited by Méndez-Vilas A. Badajoz, Spain: Formatex; 2007:758-765.
• [9]Pammi M, Flores A, Leeflang M, Versalovic J: Molecular assays in the diagnosis of neonatal sepsis: a systematic review and meta-analysis. Pediatrics 2011, 128(4):e973-e985.
• [10]Venkatesh M, Flores A, Luna RA, Versalovic J: Molecular microbiological methods in the diagnosis of neonatal sepsis. Expert Rev Anti Infect Ther 2010, 8(9):1037-1048.
• [11]Furet JP, Quenee P, Tailliez P: Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR. Int J Food Microbiol 2004, 97(2):197-207.
• [12]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(10):4506-4512.
• [13]Ventura M, Reniero R, Zink R: Specific identification and targeted characterization of Bifidobacterium lactis from different environmental isolates by a combined multiplex-PCR approach. Appl Environ Microbiol 2001, 67(6):2760-2765.
• [14]Turroni F, Peano C, Pass DA, Foroni E, Severgnini M, Claesson MJ, Kerr C, Hourihane J, Murray D, Fuligni F, et al.: Diversity of bifidobacteria within the infant guts micro biota. PLoS One 2012, 7(5):e36957.
• [15]Garland S, Tobin J, Pirotta M, Tabrizi S, Opie G, Donath S, Tang M, Morley C, Hickey L, Ung L, et al.: The ProPrems trial: investigating the effects of probiotics on late onset sepsis in very preterm infants. BMC Infect Dis 2011, 11(1):210. BioMed Central Full Text