The analysis of microbial communities is of increasing importance in life sciences and bioengineering. Traditional techniques of investigations like culture or cloning methods suffer from many disadvantages. They are unable to give a complete qualitative and quantitative view of the total amount of microorganisms themselves, their interactions among each other and with their environment. Obviously, the determination of static or dynamic balances among microorganisms is of fast growing interest. The generation of species specific and fluorescently labeled 16S ribosomal DNA (rDNA) fragments by the terminal restriction fragment length polymorphism (T-RFLP) technique is a suitable tool to overcome the problems other methods have. For the separation of these fragments polyacrylamide gel sequencers are preferred as compared to capillary sequencers using linear polymers until now because of their higher electrophoretic resolution and therefore sizing accuracy. But modern capillary sequencers, especially multicapillary sequencers, offer an advanced grade of automation and an increased throughput necessary for the investigation of complex communities in long-time studies. Therefore, we adapted a T-RFLP technique to an automated high-throughput multicapillary electrophoresis device (ABI 3100 Genetic Analysis) with regard to a precise qualitative and quantitative characterization of microbial communities.
We consider the rapid ribosequencing method as a promising new tool for the analysis of infectious agents in primarily sterile body fluids where conventional culturing of microorganisms fails.
A deep absceding infection is reported of the inframandibular part of the face of a 22-year-old male student due to a zoophilic strain of Trichophyton interdigitale. The fungus was probably acquired from the cat of the patient. Initial therapy by a general practitioner was with topical glucocorticoids and oral antihistaminica. The patient developed a severe phlegmoneous inflammation of the bearded part of the face. Later, the patient was successfully treated by a combination of itraconazole and fluconazole. Identification of the species was confirmed by light and scanning microscopy as well as sequence analysis of the internal transcribed spacer region of the ribosomal DNA.
Until now, Blastobacter denitrificans has not been mentioned in the context of human infections. A case of severe complication caused by B. denitrificans after routine appendectomy in a young girl is described and confirms this organism to be an opportunistic human pathogen
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