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Bacteriophage diagnostic

Jordi Rello, Eleonora Bunsow, Antonio Perez
Bacterial resistance to antibiotics is increasing worldwide, due to the emergence of multidrug-resistant strains. With this panorama, there is a serious danger that we may be entering the 'post-antibiotic era'. Areas covered: We assess why so few new classes of antibiotics have been developed in the past years and discuss a variety of treatments that may be able to replace antimicrobials: monoclonal antibodies, bacteriophages, stem cells and anti-virulence agents such as liposomes. Expert commentary: There are a series of economic, scientific-research and regulatory reasons for the scarcity of new antimicrobials...
October 6, 2016: Expert Review of Clinical Pharmacology
Ioana Laura Aanei, Adel M ElSohly, Michelle E Farkas, Chawita Netirojjanakul, Melanie Regan, Stephanie Taylor Murphy, James P O'Neil, Youngho Seo, Matthew B Francis
A variety of nanoscale scaffolds, including virus-like particles (VLPs), are being developed for biomedical applications; however, little information is available about their in vivo behavior. Targeted nanoparticles are particularly valuable as diagnostic and therapeutic carriers because they can increase the signal-to-background ratio of imaging agents, improve the efficacy of drugs, and reduce adverse effects by concentrating the therapeutic molecule in the region of interest. The genome-free capsid of bacteriophage MS2 has several features that make it well-suited for use in delivery applications, such as facile production and modification, the ability to display multiple copies of targeting ligands, and the capacity to deliver large payloads...
September 9, 2016: Molecular Pharmaceutics
Maryury Brown-Jaque, Maite Muniesa, Ferran Navarro
Bacteriophages are viruses that infect bacteria, and they are found everywhere their bacterial hosts are present, including the human body. To explore the presence of phages in clinical samples, we assessed 65 clinical samples (blood, ascitic fluid, urine, cerebrospinal fluid, and serum). Infectious tailed phages were detected in >45% of ascitic fluid and urine samples. Three examples of phage interference with bacterial isolation were observed. Phages prevented the confluent bacterial growth required for an antibiogram assay when the inoculum was taken from an agar plate containing lysis plaques, but not when taken from a single colony in a phage-free area...
2016: Scientific Reports
Valery A Petrenko, James W Gillespie
INTRODUCTION: New phage-directed nanomedicines have emerged recently as a result of the in-depth study of the genetics and structure of filamentous phage and evolution of phage display and phage nanobiotechnology. This review focuses on the progress made in the development of the cancer-targeted nanomaterials and discusses the trends in using phage as a bioselectable molecular navigation system. AREAS COVERED: The merging of phage display technologies with nanotechnology in recent years has proved promising in different areas of medicine and technology, such as medical diagnostics, molecular imaging, vaccine development and targeted drug/gene delivery, which is the focus of this review...
July 28, 2016: Expert Opinion on Drug Delivery
Carlos G Leon-Velarde, Lotta Happonen, Maria Pajunen, Katarzyna Leskinen, Andrew M Kropinski, Laura Mattinen, Monika Rajtor, Joanna Zur, Darren Smith, Shu Chen, Ayesha Nawaz, Roger P Johnson, Joseph A Odumeru, Mansel W Griffiths, Mikael Skurnik
UNLABELLED: Bacteriophages present huge potential both as a resource for developing novel tools for bacterial diagnostics and for use in phage therapy. This potential is also valid for bacteriophages specific for Yersinia enterocolitica To increase our knowledge of Y. enterocolitica-specific phages, we characterized two novel yersiniophages. The genomes of the bacteriophages vB_YenM_TG1 (TG1) and vB_YenM_ϕR1-RT (ϕR1-RT), isolated from pig manure in Canada and from sewage in Finland, consist of linear double-stranded DNA of 162,101 and 168,809 bp, respectively...
September 1, 2016: Applied and Environmental Microbiology
Joey N Talbert, Samuel D Alcaine, Sam R Nugen
Bacteriophages represent multifaceted building blocks that can be incorporated as substitutes for, or in unison with other detection methods, to create powerful new diagnostics for the detection of bacteria. The ease of phage manipulation, production, and detection speed clearly highlights that there remains unrealized opportunities to leverage these phage-based components in diagnostics amenable to resource-limited settings. The passage of regulations like the Food Safety Modernization act, and the ever increasing extent of global trade and travel, will create further demand for these types of diagnostics...
April 2016: Bioengineered
N R Telesmanich, E V Goncharenko, S O Chaika, I A Chaika, V O Telicheva
AIM: Study mechanisms of interaction of diagnostic bacteriophage El Tor with sensitive strain Vibrio cholerae El Tor 18507 using direct protein profiling, identification of constant and variable proteins, taking part in interaction of the phage and cell, as well as carbohydrate-specific phage receptors. MATERIALS AND METHODS: . A commercial preparation of cholera diagnostic bacteriophage El Tor, strain V. cholerae El Tor 18507 were used. Effect of carbohydrates on bacteriophage activity was determined in experiments with phage by a classic and modified by us method...
March 2016: Zhurnal Mikrobiologii, Epidemiologii, i Immunobiologii
Anupam A Sawant, Progya P Mukherjee, Rahul K Jangid, Sanjeev Galande, Seergazhi G Srivatsan
The development of robust tools and practical RNA labeling strategies that would facilitate the biophysical analysis of RNA in both cell-free and cellular systems will have profound implications in the discovery of new RNA diagnostic tools and therapeutic strategies. In this context, we describe the development of a new alkyne-modified UTP analog, 5-(1,7-octadinyl)uridine triphosphate (ODUTP), which serves as an efficient substrate for the introduction of a clickable alkyne label into RNA transcripts by bacteriophage T7 RNA polymerase and mammalian cellular RNA polymerases...
June 28, 2016: Organic & Biomolecular Chemistry
Svetlana P Ikonomova, Ziming He, Amy J Karlsson
Antibody fragments, such as the single-chain variable fragment (scFv), have much potential in research and diagnostics because of their antigen-binding ability similar to a full-sized antibody and their ease of production in microorganisms. Some applications of antibody fragments require immobilization on a surface, and we have established a simple immobilization method that is based on the biotin-streptavidin interaction and does not require a separate purification step. We genetically fused two biotinylation tags-the biotin carboxyl carrier protein (BCCP) or the AviTag minimal sequence-to six different scFvs (scFv13R4, scFvD10, scFv26-10, scFv3, scFv5, and scFv12) for site-specific biotinylation in vivo by endogenous biotin ligases produced by Escherichia coli...
August 2016: Journal of Immunological Methods
Dana Braff, David Shis, James J Collins
The growing prevalence of antibiotic resistance calls for new approaches in the development of antimicrobial therapeutics. Likewise, improved diagnostic measures are essential in guiding the application of targeted therapies and preventing the evolution of therapeutic resistance. Discovery platforms are also needed to form new treatment strategies and identify novel antimicrobial agents. By applying engineering principles to molecular biology, synthetic biologists have developed platforms that improve upon, supplement, and will perhaps supplant traditional broad-spectrum antibiotics...
April 18, 2016: Advanced Drug Delivery Reviews
Ruqaiyyah Siddiqui, Huma Kulsoom, Salima Lalani, Naveed Ahmed Khan
Balamuthia mandrillaris is a protist pathogen that can cause encephalitis with a mortality rate of more than 95%. Early diagnosis followed by aggressive treatment is a pre-requisite for successful prognosis. Current methods for identifying this organism rely on culture and microscopy, antibody-based methods using animals, or involve the use of molecular tools that are expensive. Here, we describe the isolation of antibody fragments that can be used for the unequivocal identification of B. mandrillaris. B. mandrillaris-specific antibody fragments were isolated from a bacteriophage antibody display library...
July 2016: Experimental Parasitology
S D Alcaine, K Law, S Ho, A J Kinchla, D A Sela, S R Nugen
Bacteriophage (phage) amplification is an attractive method for the detection of bacteria due to a narrow phage-host specificity, short amplification times, and the phages' ability to differentiate between viable and non-viable bacterial cells. The next step in phage-based bacteria detection is leveraging bioengineered phages to create low-cost, rapid, and easy-to-use detection platforms such as lateral flow assays. Our work establishes the proof-of-concept for the use of bioengineered T7 phage strains to increase the sensitivity of phage amplification-based lateral flow assays...
August 15, 2016: Biosensors & Bioelectronics
A A Byvalov, L G Dudina, I V Konyshev, S G Litvinets, E A Martinson
The effect of treatment of Yersinia pseudotuberculosis cells with antibodies of various specificities on adhesiveness of pseudotuberculosis bacteriophage was analyzed by competitive inhibition technique. Bacteriophage adsorption to bacteria was sterically inhibited by monoclonal antibodies to protein epitopes of Y. pseudotuberculosis outer membrane. These results suggest that receptors of pseudotuberculosis diagnostic bacteriophage are localized on the LPS core of microbial cell.
March 2016: Bulletin of Experimental Biology and Medicine
Cameron S Ball, Yooli K Light, Chung-Yan Koh, Sarah S Wheeler, Lark L Coffey, Robert J Meagher
Reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) has frequently been proposed as an enabling technology for simplified diagnostic tests for RNA viruses. However, common detection techniques used for LAMP and RT-LAMP have drawbacks, including poor discrimination capability, inability to multiplex targets, high rates of false positives, and (in some cases) the requirement of opening reaction tubes postamplification. Here, we present a simple technique that allows closed-tube, target-specific detection, based on inclusion of a dye-labeled primer that is incorporated into a target-specific amplicon if the target is present...
April 5, 2016: Analytical Chemistry
Priscillia Lagoutte, Charlotte Mignon, Stéphanie Donnat, Gustavo Stadthagen, Jan Mast, Régis Sodoyer, Adrien Lugari, Bettina Werle
Virus-like particles (VLPs) are promising molecular structures for the design and construction of novel vaccines, diagnostic tools, and gene therapy vectors. Size, oligomer assembly and repetitiveness of epitopes are optimal features to induce strong immune responses. Several VLP-based vaccines are currently licensed and commercialized, and many vaccine candidates are now under preclinical and clinical studies. In recent years, the development of genetically engineered recombinant VLPs has accelerated the need for new, improved downstream processes...
June 2016: Journal of Virological Methods
Joseph R Merrill, Krzysztof Krajewski, Hong Yuan, Jonathan E Frank, David S Lalush, Cam Patterson, Anka N Veleva
Tumor angiogenesis, the formation of new tumor blood supply, has been recognized as a hallmark of cancer and represents an important target for clinical management of various angiogenesis-dependent solid tumors. Previously, by screening a bacteriophage peptide library we have discovered the FHT-peptide sequence that binds specifically to bone marrow-derived tumor vasculature with high affinity. Here in an effort to determine the potential of the FHT-peptide for in vivo positron emission tomography (PET) imaging of aggressive tumor vasculature we studied four FHT-derivatives: NOTA-FHT, NOTA-(FHT)2, NOTA-PEG-FHT, and NOTA-PEG-(FHT)2...
April 2016: Biomaterials
Louise E Britton, Joseph P Cassidy, Jim O'Donovan, Stephen V Gordon, Bryan Markey
Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease (paratuberculosis), a chronic wasting disease in cattle with important welfare, economic and potential public health implications. Current tests are unable to recognise all stages of the disease, which makes it difficult to diagnose and control. This review explores emerging diagnostic techniques that could complement and enhance the diagnosis of MAP infection, including bacteriophage analysis, new MAP-specific antigens, host protein expression in response to infection, transcriptomic studies, analysis of microRNAs and investigation of the gastrointestinal microbiome...
March 2016: Veterinary Journal
David J Simpson, Jessica C Sacher, Christine M Szymanski
Recently, a large number of new technologies have been developed that exploit the unique properties of bacteriophage receptor binding proteins (RBPs). These include their use in diagnostic applications that selectively capture bacteria and as therapeutics that reduce bacterial colonization in vivo. RBPs exhibit comparable, and in many cases superior, stability, receptor specificity, and affinity to other carbohydrate binding proteins such as antibodies or lectins. In order to further exploit the use of RBPs, we have developed an assay for discovering RBPs using phage genome expression libraries and protein screens to identify binding partners that recognize the host bacterium...
January 2016: Viruses
Christopher G Hosking, Hamish E G McWilliam, Patrick Driguez, David Piedrafita, Yuesheng Li, Donald P McManus, Leodevico L Ilag, Els N T Meeusen, Michael J de Veer
The development of effective diagnostic tools will be essential in the continuing fight to reduce schistosome infection; however, the diagnostic tests available to date are generally laborious and difficult to implement in current parasite control strategies. We generated a series of single-chain antibody Fv domain (scFv) phage display libraries from the portal lymph node of field exposed water buffaloes, Bubalus bubalis, 11-12 days post challenge with Schistosoma japonicum cercariae. The selected scFv-phages showed clear enrichment towards adult schistosomes and excretory-secretory (ES) proteins by immunofluorescence, ELISA and western blot analysis...
December 2015: PLoS Neglected Tropical Diseases
Ashwani Sharma, Farzin Haque, Fengmei Pi, Lyudmila S Shlyakhtenko, B Mark Evers, Peixuan Guo
UNLABELLED: We report programmable self-assembly of branched, 3D globular, monodisperse and nanoscale sized dendrimers using RNA as building blocks. The central core and repeating units of the RNA dendrimer are derivatives of the ultrastable three-way junction (3WJ) motif from the bacteriophage phi29 motor pRNA. RNA dendrimers were constructed by step-wise self-assembly of modular 3WJ building blocks initiating with a single 3WJ core (Generation-0) with overhanging sticky end and proceeding in a radial manner in layers up to Generation-4...
April 2016: Nanomedicine: Nanotechnology, Biology, and Medicine
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