A methodology for using Lambda phages as a proxy for pathogen transmission in hospitals

Burke, Kylie B; Berryhill, Brandon A; García, Rodrigo Villalobos; Goldberg, David; Manuel, Joshua A; Gannon, Paige; Levin, Bruce R.; Kraft, Colleen S.; Mumma, Joel M.

doi:10.1016/j.jhin.2023.01.004

Cited by 4 publications

(7 citation statements)

References 12 publications

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“…Using a previously validated procedure, 4 we sprayed with small spray bottles (Bürkle, Germany, #10216-888) with high densities (approximately 10 8 phages per mL) of three genetically distinct λ phages on HCP volunteers to reflect densities of pathogens found in patient samples (Supplementary Table 2). 2 mL of λ phages were sprayed onto three locations, and each site was sprayed with a different variant of λ: one marked with a kanamycin resistance gene was sprayed on the wrists; one marked with a chloramphenicol resistance gene was sprayed on the back of the hood; and one lacking an antibiotic marker at the critical triangle, described below.…”

Section: Phage Testingmentioning

confidence: 99%

“…Each type of λ phage lysate was inoculated, shaken, centrifuged, and filtered per the methods in Burke et al 4 to create high-titer lysates (PFU/mL between 1×10 8 and 1×10 9 ). These lysates were stored in spray bottles, transported, and primed per the methods in Burke et al 4 Immediately after priming, each lysate was sprayed with one pump from a distance of 10 cm onto the target sites for initial contamination. The spray dried clear and was unidentifiable to the naked eye.…”

Section: Bacteriophage Lysate Preparation and Distributionmentioning

confidence: 99%

“…Phage identification was performed by polymerase chain reaction (PCR), using the methods and materials used by Burke et al 4 Band sizes of 800bp were called λ Temp , 1500bp called λ Chl , and 1900bp called λ Kan . The PCR was performed with an O'Gene Ruler DNA Ladder (Thermo Fisher Scientific, USA, #SM1563).…”

Section: Bacteriophage Identification and Quantificationmentioning

confidence: 99%

“…The viruses are visually undetectable and can be inactivated via alcohol-based sanitation but pose no appreciable risk. 4,14 With the original doffing protocol, all participants had at least one contamination with at least 1800 virions present-an amount far greater than the minimum infective dose of many HCIDs (Supplementary Table 2).…”

Section: Event Descriptionmentioning

confidence: 99%

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A bacteriophage-based validation of a personal protective equipment doffing procedure to be used with high-consequence pathogens

Berryhill,

Burke,

Smith

et al. 2024

Infect. Control Hosp. Epidemiol.

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Objective: To determine if the high-level personal protective equipment used in the treatment of high-consequence infectious diseases is effective at stopping the spread of pathogens to healthcare personnel (HCP) while doffing. Background: Personal protective equipment (PPE) is fundamental to the safety of HCPs. HCPs treating patients with high-consequence infectious diseases use several layers of PPE, forming complex protective ensembles. With high-containment PPE, step-by-step procedures are often used for donning and doffing to minimize contamination risk to the HCP, but these procedures are rarely empirically validated and instead rely on following infection prevention best practices. Methods: A doffing protocol video for a high-containment PPE ensemble was evaluated to determine potential contamination pathways. These potential pathways were tested using fluorescence and genetically marked bacteriophages. Results: The experiments revealed existing protocols permit contamination pathways allowing for transmission of bacteriophages to HCPs. Updates to the doffing protocols were generated based on the discovered contamination pathways. This updated doffing protocol eliminated the movement of viable bacteriophages from the outside of the PPE to the skin of the HCP. Conclusions: Our results illustrate the need for quantitative, scientific investigations of infection prevention practices, such as doffing PPE.

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Section: Phage Testingmentioning

confidence: 99%

Section: Bacteriophage Lysate Preparation and Distributionmentioning

confidence: 99%

Section: Bacteriophage Identification and Quantificationmentioning

confidence: 99%

Section: Event Descriptionmentioning

confidence: 99%

See 2 more Smart Citations

A bacteriophage-based validation of a personal protective equipment doffing procedure to be used with high-consequence pathogens

Berryhill,

Burke,

Smith

et al. 2024

Infect. Control Hosp. Epidemiol.

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“…Participants knew that contamination may be present in the simulation but were unaware of the location of contamination and sampling sites. For more detail about our viral surrogate methodology, see Burke et al (2023).…”

Section: Measuring Contamination Performancementioning

confidence: 99%

Sequencing Patient Care Tasks as an Infection Prevention Practice

Weaver,

Mumma,

Parmar

et al. 2023

Proceedings of the Human Factors and Ergonomics Society Annual

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Healthcare-associated infections are common, yet largely preventable. We examined whether nurses that successfully limited their contamination spread in a high-fidelity simulated environment sequenced their tasks differently than nurses that spread more contamination by reanalyzing an existing dataset. In the simulations, contamination spread was tracked using live viral surrogates (bacteriophages), which are harmless to humans. An overall contamination performance score was calculated for each participant, who were divided into one of three performance groups: high (M = 93%), medium (M = 78%), or low (M = 59%). An ANOVA showed contamination performance group did not have a statistically significant effect on the order nurses completed tasks; the largest effect size was small ( ηG2 = 0.019). Thus, even if nurses that successfully limit their contamination spread do sequence their tasks differently, it may not be practically meaningful because it is a small effect.

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Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures

Pfuderer

Stark

Grass

2023

ACS Appl. Bio Mater.

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Effective cleaning and disinfection procedures are an integral part of good manufacturing practice and in maintaining hygiene standards in health-care facilities. In this study, a method to validate such cleaning and disinfection procedures of surfaces was established employing lipid nanoparticles (LNPs) encapsulating DNA. It was possible to determine and distinguish between the physical cleaning effect (dilution) and the chemical cleaning effect (disintegration) on the LNPs during the cleaning and disinfection procedure (wiping). After treatment with 70 v % ethanol as a disinfectant and SDS solution as a cleaning agent, LNPs showed log 10 reductions of 4.5 and 4.0, respectively. These values are similar to the log 10 reductions exhibited by common bacteria, such as Escherichia coli and Serratia marcescens. Therefore, LNPs pose as useful tools for cleaning validation with advantages over the already existing tools and enable a separate detection of dilution and chemical disinfectant action.

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A methodology for using Lambda phages as a proxy for pathogen transmission in hospitals

Cited by 4 publications

References 12 publications

A bacteriophage-based validation of a personal protective equipment doffing procedure to be used with high-consequence pathogens

A bacteriophage-based validation of a personal protective equipment doffing procedure to be used with high-consequence pathogens

Sequencing Patient Care Tasks as an Infection Prevention Practice

Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures

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