Comparison of anti-fouling surface coatings for applications in bacteremia diagnostics

Boardman, Anna K.; Allison, Sandra; Sharon, André; Sauer-Budge, Alexis F.

doi:10.1039/c2ay25662b

Cited by 6 publications

(7 citation statements)

References 47 publications

(60 reference statements)

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“…Prior to experiments, all device components were sterilized with 10% bleach and rinsed with sterile water. The molded funnels were then submerged in a beaker of 0.05% (w/v) Pluronic F-127 (Sigma-Aldrich) to minimize microbial adherence during the procedure. , The filled beaker was placed in a sonic bath for 10 min to facilitate adsorption of the Pluronic onto the parts’ surfaces.…”

Section: Experimental Sectionmentioning

confidence: 99%

Rapid Detection of Bacteria from Blood with Surface-Enhanced Raman Spectroscopy

et al. 2016

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Traditional methods for identifying pathogens in bacteremic patients are slow (24–48+ h). This can lead to physicians making treatment decisions based on an incomplete diagnosis and potentially increasing the patient’s mortality risk. To decrease time to diagnosis, we have developed a novel technology that can recover viable bacteria directly from whole blood and identify them in less than 7 h. Our technology combines a sample preparation process with surface-enhanced Raman spectroscopy (SERS). The sample preparation process enriches viable microorganisms from 10 mL of whole blood into a 200 μL aliquot. After a short incubation period, SERS is used to identify the microorganisms. We further demonstrated that SERS can be used as a broad detection method, as it identified a model set of 17 clinical blood culture isolates and microbial reference strains with 100% identification agreement. By applying the integrated technology of sample preparation and SERS to spiked whole blood samples, we were able to correctly identify both Staphylococcus aureus and Escherichia coli 97% of the time with 97% specificity and 88% sensitivity.

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Section: Experimental Sectionmentioning

confidence: 99%

Rapid Detection of Bacteria from Blood with Surface-Enhanced Raman Spectroscopy

et al. 2016

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“…The brush conformation of PEO yields non-adhesive properties due to its highly hydrated polymer chains. Previous experiments have shown that Pluronic F-127 treatment reduces bacterial adhesion to plastic surfaces [ 37 ].…”

Section: Resultsmentioning

confidence: 99%

“…Prior to experiments, all device components were sterilized with 10% bleach and rinsed with sterile water. The molded funnels were then submerged in a beaker of 0.05% (w/v) Pluronic F-127 (Sigma Aldrich) to minimize microbial adherence during the procedure [ 37 ]. This beaker was placed in a sonic bath for 10 min to help facilitate adsorption of the Pluronic onto the parts’ surfaces.…”

Section: Methodsmentioning

confidence: 99%

Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

et al. 2015

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Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample.

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“…A recurring problem is the significant fraction of bacteria that is accidentally lost by non-specific binding in miniaturized devices and microfluidics due to their high surface area to volume ratios. Thus, antifouling properties are of greater importance [ 155 ]. Furthermore, fouling by non-specific biomolecules can also hinder sensitivity and selectivity and result in false-positive or -negative readings [ 156 ].…”

Section: Clinical–diagnostic Application Of Pathogen Adhesion Testsmentioning

confidence: 99%

Host-Pathogen Adhesion as the Basis of Innovative Diagnostics for Emerging Pathogens

Belkum¹,

Almeida²,

Bardiaux

et al. 2021

Diagnostics

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Infectious diseases are an existential health threat, potentiated by emerging and re-emerging viruses and increasing bacterial antibiotic resistance. Targeted treatment of infectious diseases requires precision diagnostics, especially in cases where broad-range therapeutics such as antibiotics fail. There is thus an increasing need for new approaches to develop sensitive and specific in vitro diagnostic (IVD) tests. Basic science and translational research are needed to identify key microbial molecules as diagnostic targets, to identify relevant host counterparts, and to use this knowledge in developing or improving IVD. In this regard, an overlooked feature is the capacity of pathogens to adhere specifically to host cells and tissues. The molecular entities relevant for pathogen–surface interaction are the so-called adhesins. Adhesins vary from protein compounds to (poly-)saccharides or lipid structures that interact with eukaryotic host cell matrix molecules and receptors. Such interactions co-define the specificity and sensitivity of a diagnostic test. Currently, adhesin-receptor binding is typically used in the pre-analytical phase of IVD tests, focusing on pathogen enrichment. Further exploration of adhesin–ligand interaction, supported by present high-throughput “omics” technologies, might stimulate a new generation of broadly applicable pathogen detection and characterization tools. This review describes recent results of novel structure-defining technologies allowing for detailed molecular analysis of adhesins, their receptors and complexes. Since the host ligands evolve slowly, the corresponding adhesin interaction is under selective pressure to maintain a constant receptor binding domain. IVD should exploit such conserved binding sites and, in particular, use the human ligand to enrich the pathogen. We provide an inventory of methods based on adhesion factors and pathogen attachment mechanisms, which can also be of relevance to currently emerging pathogens, including SARS-CoV-2, the causative agent of COVID-19.

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Comparison of anti-fouling surface coatings for applications in bacteremia diagnostics

Cited by 6 publications

References 47 publications

Rapid Detection of Bacteria from Blood with Surface-Enhanced Raman Spectroscopy

Rapid Detection of Bacteria from Blood with Surface-Enhanced Raman Spectroscopy

Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

Host-Pathogen Adhesion as the Basis of Innovative Diagnostics for Emerging Pathogens

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