Bacteriophage-Based Pathogen Detection

Ripp, Steven

doi:10.1007/10_2009_7

Cited by 20 publications

(12 citation statements)

References 52 publications

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“…The neutralizer test also was done as described in the above-cited study 36 . Briefly, after 5 min of contact time with YPP-100 and removing the excess YPP-100 as described above, each of the phage-treated matrices was mixed (30 sec) gently in separate conical tubes (50-mL capacity) containing peptone water (20 mL), the mixtures were passed through membrane filters (0.45-µm pore-size) and the filters were washed with PBS (20 mL).…”

Section: Methodsmentioning

confidence: 99%

A Yersinia pestis-specific, lytic phage preparation significantly reduces viable Y. pestis on various hard surfaces experimentally contaminated with the bacterium

et al. 2012

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Five Y. pestis bacteriophages obtained from various sources were characterized to determine their biological properties, including their taxonomic classification, host range and genomic diversity. Four of the phages (YpP-G, Y, R and YpsP-G) belong to the Podoviridae family, and the fifth phage (YpsP-PST) belongs to the Myoviridae family, of the order Caudovirales comprising of double-stranded DNA phages. The genomes of the four Podoviridae phages were fully sequenced and found to be almost identical to each other and to those of two previously characterized Y. pestis phages Yepe2 and φA1122. However, despite their genomic homogeneity, they varied in their ability to lyse Y. pestis and Y. pseudotuberculosis strains. The five phages were combined to yield a “phage cocktail” (tentatively designated “YPP-100”) capable of lysing the 59 Y. pestis strains in our collection. YPP-100 was examined for its ability to decontaminate three different hard surfaces (glass, gypsum board and stainless steel) experimentally contaminated with a mixture of three genetically diverse Y. pestis strains CO92, KIM and 1670G. Five minutes of exposure to YPP-100 preparations containing phage concentrations of ca. 109, 108 and 107 PFU/mL completely eliminated all viable Y. pestis cells from all three surfaces, but a few viable cells were recovered from the stainless steel coupons treated with YPP-100 diluted to contain ca. 106 PFU/mL. However, even that highly diluted preparation significantly (p = < 0.05) reduced Y. pestis levels by ≥ 99.97%. Our data support the idea that Y. pestis phages may be useful for decontaminating various hard surfaces naturally- or intentionally-contaminated with Y. pestis.

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Section: Methodsmentioning

confidence: 99%

A Yersinia pestis-specific, lytic phage preparation significantly reduces viable Y. pestis on various hard surfaces experimentally contaminated with the bacterium

et al. 2012

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“…These new assays take many forms, including direct visualization of differently labeled phages, reporter phages that genetically deposit trackable signals within their bacterial hosts, and the detection of progeny phages or other uniquely identifiable elements released from infected host cells (6,171,172,176). As far as sepsis is concerned, a phage-based assay (MicroPhage MRSA/MSSA blood culture test; Longmont) for the identification of Staphylococcus aureus and the detection of methicillin resistance directly from positive blood cultures is currently undergoing clinical trials in the United States.…”

Section: Assays For Identification Of Pathogens From Positive Blood Cmentioning

confidence: 99%

The Era of Molecular and Other Non-Culture-Based Methods in Diagnosis of Sepsis

et al. 2010

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SUMMARY Sepsis, a leading cause of morbidity and mortality throughout the world, is a clinical syndrome with signs and symptoms relating to an infectious event and the consequent important inflammatory response. From a clinical point of view, sepsis is a continuous process ranging from systemic inflammatory response syndrome (SIRS) to multiple-organ-dysfunction syndrome (MODS). Blood cultures are the current “gold standard” for diagnosis, and they are based on the detection of viable microorganisms present in blood. However, on some occasions, blood cultures have intrinsic limitations in terms of sensitivity and rapidity, and it is not expected that these drawbacks will be overcome by significant improvements in the near future. For these principal reasons, other approaches are therefore needed in association with blood culture to improve the overall diagnostic yield for septic patients. These considerations have represented the rationale for the development of highly sensitive and fast laboratory methods. This review addresses non-culture-based techniques for the diagnosis of sepsis, including molecular and other non-culture-based methods. In particular, the potential clinical role for the sensitive and rapid detection of bacterial and fungal DNA in the development of new diagnostic algorithms is discussed.

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“…Nevertheless, phages were originally described as therapeutic elements to treat human and animal infections (34). This application, known as phage therapy, has regained interest in the past years, particularly in an era when antibiotic resistance and biofilm-based infections are permanent issues (25). Bacteriophages are denominated "temperate" when their genetic material is integrated within the bacterial genome with no immediate lysis of the bacterium until, under certain conditions, the expression of the viral genome is induced and the production of new virus particles lyses the host cell; they are called "lytic" or "virulent" when, immediately after the infection, they redirect the bacterial metabolism to the production of new phages, which are released during the bacterial lysis (22,36).…”

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confidence: 99%

Isolation of a Novel Bacteriophage Specific for the Periodontal Pathogen Fusobacterium nucleatum

Machuca

Daille

Vinés

et al. 2010

Appl Environ Microbiol

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Fusobacterium nucleatum is a periodontal pathogen that has been directly associated with the development and progression of periodontal disease, a widespread pathology that affects the support tissues of the tooth. We isolated a new bacteriophage (Fnp⌽02) that specifically infects this bacterium. Transmission electron microscopy showed that the virion is composed of an icosahedral head and a segmented tail. The size of the phage genome was estimated to be approximately 59 kbp of double-stranded DNA. The morphological features and the genetic characteristics suggest that Fnp⌽02 is part of the Siphoviridae family. Using one-step growth and adsorption experiments, the latent period, burst size, and adsorption rate were estimated to be 15 h, 100 infectious units per cell, and 7.5 ؋ 10 ؊10 ml min ؊1 , respectively. A small fragment of phage DNA was cloned and sequenced, showing 93% nucleotide identity with the phage PA6 of Propionibacterium acnes and amino acid identity with fragments of two proteins (Gp3 and Gp4) of this phage. To our knowledge, Fnp⌽02 is the first phage described to infect Fusobacterium nucleatum and provides the base for future exploration of phages in the control of periodontal disease.

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Bacteriophage-Based Pathogen Detection

Cited by 20 publications

References 52 publications

A Yersinia pestis-specific, lytic phage preparation significantly reduces viable Y. pestis on various hard surfaces experimentally contaminated with the bacterium

A Yersinia pestis-specific, lytic phage preparation significantly reduces viable Y. pestis on various hard surfaces experimentally contaminated with the bacterium

The Era of Molecular and Other Non-Culture-Based Methods in Diagnosis of Sepsis

Isolation of a Novel Bacteriophage Specific for the Periodontal Pathogen Fusobacterium nucleatum

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