Microbial inactivation for safe and rapid diagnostics of infectious samples
Jose-Luis Sagripanti1, Birgit Hülseweh, Gudrun Grote
1Wehrwissenschaftliches Institut für Schutztechnologien ABC-Schutz, Munster, Germany.
Applied and Environmental Microbiology
|August 23, 2011
Summary
Peracetic acid and cupric ascorbate rapidly inactivate diverse pathogens, including spores and viruses. These methods preserve nucleic acids and epitopes for downstream PCR and immunoassay detection, enabling safer field sample analysis.
Area of Science:
- Microbiology
- Biosecurity
- Analytical Chemistry
Background:
- Biological threat agents require stringent safety protocols for handling and analysis.
- Existing inactivation methods often compromise the integrity of microbial nucleic acids or protein epitopes, hindering subsequent detection.
- There is a need for rapid, reliable methods to inactivate pathogens in field samples for safe analysis.
Purpose of the Study:
- To identify a rapid, simple, and effective method for complete pathogen inactivation.
- To preserve microbial nucleic acid fragments for Polymerase Chain Reaction (PCR) and protein epitopes for immunoassays.
- To enable analysis of field samples under reduced containment levels.
Main Methods:
- Evaluation of formaldehyde, hydrogen peroxide, guanidium thiocyanate, glutaraldehyde, and sodium hypochlorite for pathogen inactivation.
- Testing of peracetic acid and cupric ascorbate for inactivation of bacterial spores (Bacillus atrophaeus), vegetative bacteria (Pseudomonas aeruginosa), RNA virus (Pixuna virus), and DNA virus (vaccinia virus).
- Assessment of the impact of inactivation treatments on the suitability of samples for PCR and enzyme-linked immunosorbent assay (ELISA).
Main Results:
- Formaldehyde, hydrogen peroxide, and guanidium thiocyanate failed to inactivate high titers of spores and viruses within 30 minutes.
- Glutaraldehyde and sodium hypochlorite demonstrated high microbicidal activity but destroyed PCR and ELISA detectability.
- Peracetic acid and cupric ascorbate achieved >6 log(10) inactivation of diverse pathogens within 30 minutes at 21°C with minimal impact on PCR and immunoassay detection.
Conclusions:
- Peracetic acid and cupric ascorbate are effective agents for rapid and complete inactivation of a broad spectrum of pathogens.
- These agents preserve critical biomolecules (nucleic acids, epitopes) essential for molecular and immunological detection methods.
- The findings support the development of protocols for safer, cost-effective field sample processing and analysis under lower containment levels.
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