Rapid and Biologically Safe Procedures for the Evaluation of Antigen-Specific           T Cell Response to Microbial Pathogens that May be Used in the BSL-3 and BSL-4           Environment

Agrati, Chiara; Volpi, I; Martini, Federico; Gioia, Cristiana; Castilletti, Concetta; Ippolito, Giuseppe; Capobianchi, Maria Rosaria; Poccia, Fabrizio

doi:10.1177/153567600801300105

Cited by 3 publications

(2 citation statements)

References 9 publications

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“…Moreover, among these inactivation methods, only PFA treatment is compatible with any of the potential downstream analyses of infected cells. There is one published report that described inactivation of Ft-infected cells using 3-8% PFA treatment for 30 min (Agrati et al, 2008). However, this study does not contain any primary data related to the efficacy of PFA inactivation, but instead, only cites guidance from a 2006 CDC website report that is no longer available.…”

Section: Discussionmentioning

confidence: 99%

Efficient inactivation ofBurkholderia pseudomalleiorFrancisella tularensisin infected cells for safe removal from biosafety level 3 containment laboratories

2014

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Working with infectious agents that require BSL-3 level containment agents offers many challenges for researchers. BSL-3 containment laboratories are usually not equipped with expensive specialty equipment that is needed for studies such as flow cytometric analysis, microscopy, and proteomic analyses. Therefore, for most researchers that are working with BSL-3 level infectious agents, removal of samples from BSL-3 labs for these types of studies is necessary, and methods for complete and dependable inactivation of the samples are required. In this report we have done a thorough characterization of the effectiveness of paraformaldehyde fixation for inactivation of cell samples infected with the intracellular bacterial agents Burkholderia pseudomallei (Bp) and Francisella tularensis (Ft), both of which are Tier 1 select agent pathogens that require BSL-3 containment. We have demonstrated that cells infected with these pathogens are completely inactivated via 5-minute treatment with 4% paraformaldehyde. Moreover, a 15-minute treatment with 2% paraformaldehyde completely sterilized both Bp- and Ft-infected cells. These studies also revealed that Bp is significantly more sensitive to paraformaldehyde treatment than Ft. Our findings have clearly demonstrated that a 15-minute treatment of Bp- or Ft-infected cells with 4% paraformaldehyde solution will allow for safe removal of the cell samples from BSL-3 labs for downstream studies.

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Section: Discussionmentioning

confidence: 99%

Efficient inactivation ofBurkholderia pseudomalleiorFrancisella tularensisin infected cells for safe removal from biosafety level 3 containment laboratories

2014

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show abstract

“…Moreover, among these inactivation methods, only PFA treatment is compatible with any of the potential downstream analyses of infected cells. There is one published report that described inactivation of Ft-infected cells using 3-8 % PFA treatment for 30 minutes [197]. However, this paper does not contain any primary data related to the efficacy of PFA inactivation, but instead only cites guidance from a 2006 CDC website report that is no longer available.…”

Section: Discussionmentioning

confidence: 99%

Genetic Control of Survival and Weight Loss during Pneumonic Burkholderia pseudomallei (Bp) Infection

Emery¹

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Cytometry in High-Containment Laboratories

Cohen,

Laux,

Douagi

2024

Methods in Molecular Biology

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Rapid and Biologically Safe Procedures for the Evaluation of Antigen-Specific T Cell Response to Microbial Pathogens that May be Used in the BSL-3 and BSL-4 Environment

Cited by 3 publications

References 9 publications

Efficient inactivation ofBurkholderia pseudomalleiorFrancisella tularensisin infected cells for safe removal from biosafety level 3 containment laboratories

Efficient inactivation ofBurkholderia pseudomalleiorFrancisella tularensisin infected cells for safe removal from biosafety level 3 containment laboratories

Genetic Control of Survival and Weight Loss during Pneumonic Burkholderia pseudomallei (Bp) Infection

Cytometry in High-Containment Laboratories

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