Rapid detection and identification with molecular methods

Abstract

A prerequisite for the biochemical and physiological investigation of microorganisms is the isolation and management of pure cultures. Nevertheless, most of the environmental microorganisms are graded as yet-not-cultivable because the nutritional requirements are unknown or they could not be isolated due to the fact that they are overgrown by faster growing microorganisms of the same habitat. In addition to plating techniques, isolation without cultivation and analysis of microbes could be performed by micromanipulation techniques or the application of optical tweezers followed by the utilization of PCR based technologies. Many different phenotypic and genotypic methods are presently being applied for microbial identification and classification. Several of these methods are suitable for the simultaneous detection on species level, like the analysis of the rRNA genes. Strain-related fingerprint techniques like RFLP-PFGE, nSAPD-, or RAPD PCR are dependent on the purity of the culture and its genomic DNA. They are not suitable for simultaneous detection. In this chapter isolation techniques and an assortment of different molecular biological methods concerning identification on the species and strain level of grape-, must-, and wine-related microorganisms are described. In general, advances in molecular biology providing new insights into the wonderfully complex conversion of grape juice to wine and the microbial ecology of winemaking. © 2009 Springer-Verlag Berlin Heidelberg. All rights reserved.