Poster De Conférence Année : 2024

Validation of disinfectants in BSL3 laboratories handling live Foot-and-Mouth-Disease Virus

Aurore Romey
Anthony Relmy
  • Fonction : Auteur
  • PersonId : 1075108
Guillaume Girault
Sandra Blaise-Boisseau
Labib Bakkali Kassimi

Résumé

Background/Introduction In laboratories handling live foot-and-mouth disease virus (FMDV), strict biosecurity measures are applied, including for the entry and exit of personnel, as well as for the decontamination of all equipment and the surfaces. Currently, the most commonly used disinfectant for FMDV is formaldehyde, which is highly toxic for humans. The objective of our study is to validate the efficacy of FMDV inactivation by alternative less toxic and easy-to-use disinfectants. Material and methods The efficacy of disinfectant products was tested using two different methods according to the French standard NF-T-72281 November 2014: (i) by spraying for laboratory surfaces, (ii) by misting for large equipment or for the whole laboratory (airborne disinfection). Efficacy tests consisted in keeping the virus in contact with the disinfectant product for a set period of time, depending on the product and the method used. Six FMDV serotypes were tested (O, A, Asia-1, SAT1, SAT2, SAT3). For the disinfection of surfaces, the virus was kept in contact with a 0.5% citric acid solution for two minutes on stainless steel supports. After contact, the virus was eluted in cell culture media and the inactivation efficiency was verified by viral isolation after three passages on sensitive cells. For airborne disinfection, decontamination was carried out using a Nocospray-type decontamination product fogger diffuser. The virus deposited on stainless steel supports was exposed in the equipment airlock to a 35% hydrogen peroxide solution for two hours. Supports were placed horizontally in the airlock. In parallel, control supports were prepared in the same way, but were not in contact with the product. After contact, the virus was eluted and diluted 1:1000 in cell culture media and virus inactivation was verified by viral isolation after three passages on cells and by large-volume titration method. The results were expressed as a reduction of the viral load determined by virus titration on cells and by an absence of virus after viral isolation. Results As 0.5% citric acid is not cytotoxic, after inactivation the recovered liquid was inoculated directly on cells for virus detection. The results of viral isolation show an absence of cytopathic effect (CPE) for the six FMDV serotypes. For the 35% hydrogen peroxide, the recovered fluid had to be diluted 1:1000 prior to isolation on cells or titration to avoid toxicity on cells. All the media treated with hydrogen peroxide showed no CPE in viral isolation or titration plates, while the positive control media showed CPE for both methods. A comparison of the test and control media shows that there was a reduction in viral titer and an absence of virus in the recovered fluid. Discussion and conclusions According to these encouraging results, the described disinfection methods could be considered as an alternative to other methods used so far based on the use of more toxic products. These methods have been validated under our laboratory conditions thus each laboratory must perform their own validation under their specific conditions.
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Dates et versions

anses-04803730 , version 1 (25-11-2024)

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  • HAL Id : anses-04803730 , version 1

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Anne-Laure Salomez, Aurore Romey, Anthony Relmy, Cindy Bernelin-Cottet, Guillaume Girault, et al.. Validation of disinfectants in BSL3 laboratories handling live Foot-and-Mouth-Disease Virus. EuFMD Open Session 2024, Oct 2024, Alcala de Henares, Spain. ⟨anses-04803730⟩
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