scholarly journals Specific Detection of Yersinia pestis Based on Receptor Binding Proteins of Phages

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 611 ◽  
Author(s):  
Friederike Born ◽  
Peter Braun ◽  
Holger C. Scholz ◽  
Gregor Grass
Keyword(s):  
Yersinia Pestis ◽  
Receptor Binding ◽  
Fluorescent Proteins ◽  
Diagnostic Methods ◽  
Tail Fiber ◽  
Yersinia Species ◽  
Recombinant Receptor ◽  
F1 Antigen ◽  
Phage Receptor ◽  
Bacterial Plaque

The highly pathogenic bacterium Yersinia pestis is the causative agent of plague, a notorious infectious zoonotic disease. When transmitted from person to person as pneumonic plague via droplets, Y. pestis is highly contagious and in most cases is fatal if left untreated. Thus, when plague is suspected, rapid diagnosis is crucial, as a serious course of the infection is only averted by early antibiotic therapy. The bacterium is easy to cultivate, accessible and has a high potential for nefarious use such as bioterrorism. Highly specific, rapid and easy-to-use confirmatory diagnostic methods are required to reliably identify the pathogen independently from PCR-based methods or F1 antigen-based immunological detection. Yersinia pestis specific phages such as L-413C and ΦA1122 are already used for detection of Y. pestis in bacterial plaque or biosensor assays. Here, we made use of the host specificities conferred by phage receptor binding (or tail fiber/spike) proteins (RBP) for developing a specific, fast and simple fluorescence-microscopy-based detection method for Y. pestis. Genes of putative RBP of phages L-413C (gpH) and ΦA1122 (gp17) were fused with those of fluorescent proteins and recombinant receptor-reporter fusion proteins were produced heterologously in Escherichia coli. When first tested on attenuated Y. pestis strain EV76, RBP-reporters bound to the bacterial cell surface. This assay could be completed within a few minutes using live or formaldehyde-inactivated cells. Specificity tests using cultures of closely related Yersinia species and several inactivated fully virulent Y. pestis strains exhibited high specificities of the RBP-reporters against Y. pestis. The L-413C RBP proved to be especially specific, as it only detected Y. pestis at all temperatures tested, whereas the RBP of ΦA1122 also bound to Y. pseudotuberculosis strains at 37 °C (but not at 28, 20 or 6 °C). Finally, the Y. pestis-specific capsule, produced when grown at 37 °C, significantly reduced binding of phage ΦA1122 RBP, whereas the capsule only slightly diminished binding of L-413C RBP.

scholarly journals Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1462
Author(s):  
Peter Braun ◽  
Nadja Rupprich ◽  
Diana Neif ◽  
Gregor Grass
Keyword(s):  
Bacillus Anthracis ◽  
Receptor Binding ◽  
Pathogen Detection ◽  
Fluorescent Proteins ◽  
Binding Protein ◽  
Host Cells ◽  
Close Relationship ◽  
Phage Receptor ◽  
Protein Assays ◽  
Receptor Binding Protein

Bacteriophage receptor binding proteins (RBPs) are employed by viruses to recognize specific surface structures on bacterial host cells. Recombinant RBPs have been utilized for detection of several pathogens, typically as fusions with reporter enzymes or fluorescent proteins. Identification of Bacillus anthracis, the etiological agent of anthrax, can be difficult because of the bacterium’s close relationship with other species of the Bacillus cereussensu lato group. Here, we facilitated the identification of B. anthracis using two implementations of enzyme-linked phage receptor binding protein assays (ELPRA). We developed a single-tube centrifugation assay simplifying the rapid analysis of suspect colonies. A second assay enables identification of suspect colonies from mixed overgrown solid (agar) media derived from the complex matrix soil. Thus, these tests identified vegetative cells of B. anthracis with little processing time and may support or confirm pathogen detection by molecular methods such as polymerase chain reaction.

Chemical composition and biological activity of the Yersinia pestis envelope substance

1980 ◽  
Vol 30 (2) ◽  
pp. 506-512
Author(s):  
R Głosnicka ◽  
E Gruszkiewicz
Keyword(s):  
Yersinia Pestis ◽  
Lipid Fraction ◽  
High Specificity ◽  
Biological Properties ◽  
Two Dimensional ◽  
Purification Procedure ◽  
Hemagglutination Activity ◽  
Passive Hemagglutination ◽  
Phage Receptor ◽  
Envelope Antigen

Purification of the envelope antigen of Yersinia pestis EV with passive hemagglutination activity is described. The purification procedure consisted of pancreatin digestion, chromatography on human erythrocyte stroma set on Celite, and rechromatography on Sephadex G-200. Chemical, physical, and biological properties of this antigen were investigated. The results show the lipid-polysaccharide structure of the isolated antigen. The carbohydrate moiety of the galactolipid antigen consists of galactose and fucose. The lipid fraction contained phosphatidylethanolamine and phosphatidylserine. The preparation showed high specificity in the hemagglutination reaction and in Y. pestis phage receptor activity. In two-dimensional immunoelectrophoresis, the isolated pancreatic envelope digest antigen appeared as a single line. Two-dimensional immunoelectrophoresis was modified for tandem separation and was employed to electrophoretically identify the pancreatic envelope digest, trypsin envelope digest preparation, and F1 envelope antigen of Y. pestis. Related or identical antigens showed confluence of peaks with reactions of identity.

scholarly journals The human PTH2 receptor: binding and signal transduction properties of the stably expressed recombinant receptor.

Endocrinology ◽  
1996 ◽  
Vol 137 (7) ◽  
pp. 2748-2757 ◽  
Author(s):  
V Behar ◽  
M Pines ◽  
C Nakamoto ◽  
Z Greenberg ◽  
A Bisello ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Receptor Binding ◽  
Recombinant Receptor

scholarly journals Exploiting phage receptor binding proteins to enable endolysins to kill Gram-negative bacteria

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Athina Zampara ◽  
Martine C. Holst Sørensen ◽  
Dennis Grimon ◽  
Fabio Antenucci ◽  
Amira Ruslanovna Vitt ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Binding Proteins ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Phage Receptor

scholarly journals Phage S144, a New Polyvalent Phage Infecting Salmonella spp. and Cronobacter sakazakii

2020 ◽  
Vol 21 (15) ◽  
pp. 5196 ◽  
Author(s):  
Michela Gambino ◽  
Anders Nørgaard Sørensen ◽  
Stephen Ahern ◽  
Georgios Smyrlis ◽  
Yilmaz Emre Gencay ◽  
...  
Keyword(s):  
Open Reading Frames ◽  
Trna Genes ◽  
Tail Fiber ◽  
Cronobacter Sakazakii ◽  
Salmonella Spp ◽  
Nucleotide Biosynthesis ◽  
C Terminus ◽  
Transmission Electron ◽  
Phage Receptor ◽  
Tail Fibers

Phages are generally considered species- or even strain-specific, yet polyvalent phages are able to infect bacteria from different genera. Here, we characterize the novel polyvalent phage S144, a member of the Loughboroughvirus genus. By screening 211 Enterobacteriaceae strains, we found that phage S144 forms plaques on specific serovars of Salmonella enterica subsp. enterica and on Cronobacter sakazakii. Analysis of phage resistant mutants suggests that the O-antigen of lipopolysaccharide is the phage receptor in both bacterial genera. The S144 genome consists of 53,628 bp and encodes 80 open reading frames (ORFs), but no tRNA genes. In total, 32 ORFs coding for structural proteins were confirmed by ESI-MS/MS analysis, whereas 45 gene products were functionally annotated within DNA metabolism, packaging, nucleotide biosynthesis and phage morphogenesis. Transmission electron microscopy showed that phage S144 is a myovirus, with a prolate head and short tail fibers. The putative S144 tail fiber structure is, overall, similar to the tail fiber of phage Mu and the C-terminus shows amino acid similarity to tail fibers of otherwise unrelated phages infecting Cronobacter. Since all phages in the Loughboroughvirus genus encode tail fibers similar to S144, we suggest that phages in this genus infect Cronobacter sakazakii and are polyvalent.

scholarly journals Molecular anatomy of the receptor binding module of a bacteriophage long tail fiber

2019 ◽  
Vol 15 (12) ◽  
pp. e1008193 ◽  
Author(s):  
Mohammad Z. Islam ◽  
Andrei Fokine ◽  
Marthandan Mahalingam ◽  
Zhihong Zhang ◽  
Carmela Garcia-Doval ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Tail Fiber ◽  
Long Tail ◽  
Molecular Anatomy

scholarly journals Flagellin Is an Effective Adjuvant for Immunization against Lethal Respiratory Challenge with Yersinia pestis

2006 ◽  
Vol 74 (2) ◽  
pp. 1113-1120 ◽  
Author(s):  
Anna N. Honko ◽  
Nammalwar Sriranganathan ◽  
Cynthia J. Lees ◽  
Steven B. Mizel
Keyword(s):  
Yersinia Pestis ◽  
Neutrophil Infiltration ◽  
Specific Ige ◽  
Innate Immune ◽  
Igg Antibody ◽  
Adjuvant Activity ◽  
Necrosis Factor Alpha ◽  
Potent Inducer ◽  
Cytokines And Chemokines ◽  
F1 Antigen

ABSTRACT Gram-negative flagellin, a Toll-like receptor 5 (TLR5) agonist, is a potent inducer of innate immune effectors such as cytokines and nitric oxide. In the lung, flagellin induces a localized and transient innate immune response characterized by neutrophil infiltration and the production of cytokines and chemokines. In view of the extraordinary potency of flagellin as an inducer of innate immunity and the contribution of innate responses to the development of adaptive immunity, we evaluated the efficacy of recombinant Salmonella flagellin as an adjuvant in an acellular plague vaccine. Mice immunized intranasally or intratracheally with the F1 antigen of Yersinia pestis and flagellin exhibited dramatic increases in anti-F1 plasma immunoglobulin G (IgG) titers that remained stable over time. In contrast, control mice had low or undetectable antibody responses. The IgG1/IgG2a ratio of antibody titers against F1 in immunized mice is consistent with a Th2 bias. However, no significant antigen-specific IgE production was detected. Interferons, tumor necrosis factor alpha, and interleukin-6 were not essential for the adjuvant effects of flagellin. Preexisting antiflagellin antibodies had no significant effect on the adjuvant activity of flagellin. Importantly, intranasal immunization with flagellin and the F1 antigen was protective against intranasal challenge with virulent Y. pestis CO92, with 93 to 100% survival of immunized mice. Lastly, vaccination of cynomolgus monkeys with flagellin and a fusion of the F1 and V antigens of Y. pestis induced a robust antigen-specific IgG antibody response.

scholarly journals Comparison of Dissociation-Enhanced Lanthanide Fluorescent Immunoassays to Enzyme-Linked Immunosorbent Assays for Detection of Staphylococcal Enterotoxin B, Yersinia pestis-Specific F1 Antigen, and Venezuelan Equine Encephalitis Virus

2001 ◽  
Vol 8 (6) ◽  
pp. 1070-1075 ◽  
Author(s):  
Darci R. Smith ◽  
Cynthia A. Rossi ◽  
Todd M. Kijek ◽  
Erik A. Henchal ◽  
George V. Ludwig
Keyword(s):  
Yersinia Pestis ◽  
Dynamic Range ◽  
Venezuelan Equine Encephalitis Virus ◽  
Staphylococcal Enterotoxin ◽  
Encephalitis Virus ◽  
Staphylococcal Enterotoxin B ◽  
Venezuelan Equine Encephalitis ◽  
Equine Encephalitis Virus ◽  
F1 Antigen ◽  
Enterotoxin B

ABSTRACT The dissociation-enhanced lanthanide fluorescent immunoassays (DELFIA) were developed for the detection of staphylococcal enterotoxin B, Yersinia pestis-specific F1 antigen, and Venezuelan equine encephalitis virus. These assays were compared to previously developed enzyme-linked immunosorbent assays (ELISAs) by determining the sensitivity or limit of detection (LOD), the dynamic range, and the reproducibility of each assay in a number of different sample matrices. The sensitivity and specificity of each assay were then determined by using a small panel of blinded spiked and nonspiked samples. All three DELFIAs demonstrated at least 1 log greater sensitivity than corresponding ELISAs utilizing the same reagents and showed an increase in dynamic range of at least 2 log10 concentrations. This increased LOD resulted in higher sensitivity rates for the DELFIA. The specificity of all of the assays evaluated was 100%, and no sample matrix effects were observed in either format. However, the reproducibility of the DELFIA was poor due to randomly distributed wells exhibiting excessive background signal (hot wells), which occurred throughout the evaluation. As this technology matures, the reproducibility of these assays should improve, as will the ability to identify hot wells. Despite its sensitivity, the logistical burden associated with the DELFIA and the technical expertise required to complete assays and interpret the data limit the application of this technology to reference or large clinical laboratories.

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