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Application of molecular biological methods for studying probiotics and the gut flora

Published online by Cambridge University Press:  09 March 2007

A. L. McCartney
A. L. McCartney*
Affiliation:
Food Microbial Sciences Unit, School of Food Biosciences, University of Reading, Whiteknights, Reading RG6 6AP, UK
*
*Corresponding author: Dr A. L. McCartney, fax +44 118 9357222, email [email protected]
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Abstract

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Increasingly, the microbiological scientific community is relying on molecular biology to define the complexity of the gut flora and to distinguish one organism from the next. This is particularly pertinent in the field of probiotics, and probiotic therapy, where identifying probiotics from the commensal flora is often warranted. Current techniques, including genetic fingerprinting, gene sequencing, oligonucleotide probes and specific primer selection, discriminate closely related bacteria with varying degrees of success. Additional molecular methods being employed to determine the constituents of complex microbiota in this area of research are community analysis, denaturing gradient gel electrophoresis (DGGE)/temperature gradient gel electrophoresis (TGGE), fluorescent in situ hybridisation (FISH) and probe grids. Certain approaches enable specific aetiological agents to be monitored, whereas others allow the effects of dietary intervention on bacterial populations to be studied. Other approaches demonstrate diversity, but may not always enable quantification of the population. At the heart of current molecular methods is sequence information gathered from culturable organisms. However, the diversity and novelty identified when applying these methods to the gut microflora demonstrates how little is known about this ecosystem. Of greater concern is the inherent bias associated with some molecular methods. As we understand more of the complexity and dynamics of this diverse microbiota we will be in a position to develop more robust molecular-based technologies to examine it. In addition to identification of the microbiota and discrimination of probiotic strains from commensal organisms, the future of molecular biology in the field of probiotics and the gut flora will, no doubt, stretch to investigations of functionality and activity of the microflora, and/or specific fractions. The quest will be to demonstrate the roles of probiotic strains in vivo and not simply their presence or absence.

Keywords

Molecular methodsProbioticsGut flora
Type
Research Article
Information
British Journal of Nutrition , Volume 88 , Issue S1 , September 2002 , pp. s29 - s37
Copyright
Copyright © The Nutrition Society 2002

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