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Detection of nucleic acid sequences from bacterial species with molecular genetic methods
E K Petershofen1, R Fislage, R Faber
1Institute of Transfusion Medicine and Immunohematology, Red Cross Blood Donor Service Center Hessia, Sandhofstrasse 1, D-60528, Frankfurt am Main, Germany.
Abstract:
While blood products become more safe in terms of viral contamination, the risk of transfusion-related bacterial infection has re-emerged to one of the major hazards in transfusion medicine. In recent prospective studies the rate of contaminated platelets ranged from 0.04 to 0.5%, and a rate of transfusion reactions between 0.007% and 0.046%. It is generally agreed that most of the organisms isolated from donated blood originate from the normal skin flora or from the environment. As it is unlikely that antiseptic methods can achieve absolute sterilization of the skin before venepuncture, blood banks have to rely on laboratory tests to detect contaminated blood products before release. But most of the currently available methods detecting bacterial contaminations do not have the potential to be sensitive and fast enough for a routine contamination screening in transfusion services. Here we present two alternative strategies based on molecular genetic techniques (Real-Time-PCR and Haystack processing) that detect or semi-quantify bacterial rRNA gene sequences for the majority of bacterial species. In addition we discuss some aspects on target selection, routine preparation and residual 16S-rDNA-contamination of enzymes.
Insights
Bacterial contamination in blood products remains a significant risk. New molecular genetic techniques like Real-Time-PCR offer faster and more sensitive detection methods for transfusion safety.
Area of Science:
- Transfusion Medicine
- Microbiology
- Molecular Biology
Background:
- Bacterial contamination of blood products is a growing concern in transfusion medicine.
- Current detection methods lack the sensitivity and speed required for routine screening.
- Donated blood contamination often stems from skin flora or environmental sources.
Purpose of the Study:
- To present novel molecular genetic strategies for detecting bacterial contamination in blood products.
- To address the limitations of existing bacterial detection methods in transfusion services.
Main Methods:
- Development and application of Real-Time-PCR for bacterial detection.
- Utilizing Haystack processing for semi-quantification of bacterial rRNA gene sequences.
- Discussion of target selection, routine preparation, and enzyme contamination.
Main Results:
- Real-Time-PCR and Haystack processing demonstrate potential for sensitive and rapid bacterial detection.
- These molecular techniques can detect or semi-quantify bacterial rRNA gene sequences.
Conclusions:
- Molecular genetic techniques offer promising alternatives for routine bacterial contamination screening in transfusion services.
- Improved detection methods are crucial for enhancing transfusion safety and mitigating transfusion-related infections.