For debate: Is disinfection of specimens, which may contain unknown or bio-terrorist organisms, essential before electron microscopic examination?

Madeley, C. R.; Biel, Stefan S.

doi:10.1016/j.jinf.2005.09.009

Cited by 7 publications

(6 citation statements)

References 6 publications

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“…Exposure of lab personnel and the public by accidentally released viruses during handling of virus–infected biological material must be avoided by efficient and feasible safety precautions. Since most electron microscopes are operated at biosafety level two and below, proper sample inactivation is necessary to guarantee biosafety [ 11 , 12 , 13 ]. For diagnostic purposes, the inactivation procedures must not only inactivate the viruses, but simultaneously preserve their morphology.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of Diagnostic Electron Microscopy

Møller

Schünadel

Nitsche

et al. 2015

Viruses

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Formaldehyde (FA) fixation of infectious samples is a well-established protocol in diagnostic electron microscopy of viruses. However, published experimental data that demonstrate virus inactivation by these fixation procedures are lacking. Usually, fixation is performed immediately before the sample preparation for microscopy. The fixation procedure should transform viruses in a non–infectious but nonetheless structurally intact form in order to allow a proper diagnosis based on morphology. FA provides an essential advantage in comparison to other disinfectants, because it preserves the ultrastructure of biological material without interfering significantly with the preparation (i.e., the negative staining) and the detection of viruses. To examine the efficiency of FA inactivation, we used Vaccinia virus, Human adenovirus and Murine norovirus as models and treated them with FA under various conditions. Critical parameters for the inactivation efficiency were the temperature, the duration of the FA treatment, and the resistance of the virus in question. Our results show that FA inactivation at low temperature (4 °C) bears a high risk of incomplete inactivation. Higher temperatures (25 °C) are more efficient, although they still require rather long incubation times to fully inactivate a complex and highly robust virus like Vaccinia. A protocol, which applied 2% buffered FA for 60 min and a temperature–shift from 25 to 37 °C after 30 min was efficient for the complete inactivation of all test viruses, and therefore has the potential to improve both biosafety and speed of diagnostic electron microscopy.

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Section: Introductionmentioning

confidence: 99%

Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of Diagnostic Electron Microscopy

Møller

Schünadel

Nitsche

et al. 2015

Viruses

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show abstract

“…1 Saccharomyces boulardii (Sb), a probiotic agent, has been shown to be effective in prevention and treatment of diarrhea due to Clostridium difficile, 2,3 enteral feeding 4 or antibiotics, 5 and traveler's diarrhea. 6 Sb is assumed to be a subtype of Saccharomyces cerevisiae (Sc) and is usually considered safe.…”

Section: Sirmentioning

confidence: 99%

Reply to: Disinfection in diagnostic electron microscopy prior to preparation?

Gelderblom

Bannert

Pauli

2007

Journal of Infection

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“…Knowledge of the detection limit and a comparison with results published for negative staining of, for instance clinical specimens, is important because samples and preparations are handled differently. Samples from a suspected bioterrorist attack are usually first inactivated by chemicals or by radiation, which may interfere with adsorption to the support (see discussion in Madeley and Biel 2006; Gelderblom et al. 2007).…”

Section: Introductionmentioning

confidence: 99%

Detection limit of negative staining electron microscopy for the diagnosis of bioterrorism-related micro-organisms

Laue

Bannert

2010

Journal of Applied Microbiology

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Aims: To determine the detection limit of diagnostic negative staining electron microscopy for the diagnosis of pathogens that could be used for bioterrorism. Methods and Results: Suspensions of vaccinia poxvirus and endospores of Bacillus subtilis were used at defined concentrations as a model for poxviruses and spores of anthrax (Bacillus anthracis), both of which are pathogens that could be used for bioterrorist attacks. Negative staining electron microscopy was performed directly or after sedimentation of these suspensions on to the sample supports using airfuge ultracentrifugation. For both virus and spores, the detection limit using direct adsorption of a 10‐μl sample volume onto the sample support was 106 particles per ml. Using airfuge ultracentrifugation with a sample volume of 80 μl, the detection limit could be reduced to 105 particles per ml for spores and to 5 × 104 particles per ml for poxviruses. The influence on particle detection of incubation time, washing and adsorption procedures was investigated. Conclusions: The reproducibility and sensitivity of the method were acceptable, particularly considering the small sample volume and low particle number applied onto the sample support. Significance and Impact of the Study: Diagnostic negative staining electron microscopy is used for the diagnosis of pathogens in emergency situations because it allows a rapid examination of all particulate matter down to the nanometre scale. This study provides precise detection limit for the method, an important factor for the validation and improvement of the technique.

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For debate: Is disinfection of specimens, which may contain unknown or bio-terrorist organisms, essential before electron microscopic examination?

Cited by 7 publications

References 6 publications

Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of Diagnostic Electron Microscopy

Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of Diagnostic Electron Microscopy

Reply to: Disinfection in diagnostic electron microscopy prior to preparation?

Detection limit of negative staining electron microscopy for the diagnosis of bioterrorism-related micro-organisms

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