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Phage of acinetobacter baumannii

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Phage of acinetobacter baumannii


The present invention provides an isolated Acinetobacter baumannii phage, comprising one or more genomic sequences selected from the group consisting of sequences of SEQ. ID. NO: 1, 2, 3 and 4, and sequences having more than 80% homology thereof. The phages of the present invention infect Acinetobacter baumannii specifically, and can be applied to reduce the amount of Acinetobacter baumannii.
Related Terms: Acinetobacter

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Inventors: Li-Kuang Chen, Nien-Tsung Lin
USPTO Applicaton #: #20120276612 - Class: 4352351 (USPTO) - 11/01/12 - Class 435 
Chemistry: Molecular Biology And Microbiology > Virus Or Bacteriophage, Except For Viral Vector Or Bacteriophage Vector; Composition Thereof; Preparation Or Purification Thereof; Production Of Viral Subunits; Media For Propagating

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The Patent Description & Claims data below is from USPTO Patent Application 20120276612, Phage of acinetobacter baumannii.

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FIELD OF INVENTION

The present invention relates to a novel phage, and more particularly, to a phage of Acinetobacter baumannii.

BACKGROUND OF THE INVENTION

Nosocomial infections are tough issues in Hospitals. Generally, the nosocomial infection rate is about from 3% to 5%. Organisms causing nosocomial infections are usually opportunistic pathogens. In other words, these bacteria are not harmful to hosts with normal immunity, and some of them are even normal flora to human; however, while hosts have weak immunity, the bacteria cause infections, resulting in diseases.

Bacteria causing nosocomial infections may exist in stethoscopes, anamnesis papers, tourniquets, grooves, syringe needles, respirators, humidifiers, furniture, floors, vents, monitors, water, soil, food (fruits, vegetables), dirt in drainage, human body such as skin, armpits, mucosal, oral cavity, upper respiratory tract, nasal cavity, gastrointestinal tract, etc.

For example, nosocomial infections occur in an intensive care unit since patients in the intensive care unit have weak immunity and have invasive therapies such as being cannulated. According to statistics, the nosocomial infection rate in an intensive care unit is about from 2% to 3%.

Currently, the most common bacteria causing nosocomial infections include Pseudomonas aeruginosa, Staphylococcus aureus, Acinetobacter baumannii, etc.

Antibiotics are general therapeutic agents for treating bacterial infections. However, when an antibiotic is overused, bacteria will be selected to have resistance to more antibiotics. In current nosocomial infections, there are more and more bacteria having resistance to antibiotics, and patients infected by these bacteria have to be treated with expensive and novel antibiotics. Further, if the antibiotic resistance keeps developed, there will be no effective antibiotic for therapy.

Acinetobacter baumannii (abbreviated as AB, hereafter) belong to Gram negative bacteria. Generally, Acinetobacter baumannii exist in skin, respiratory tract, and gastrointestinal tract in 10% population of human. Acinetobacter baumannii favor warm and humid environment, so as to exist in medical devices, water troughs, beds, bed mats, respiratory devices and even air in a hospital. Currently, Acinetobacter baumannii having multiple resistances to gentamicin, amikacin piperacillin/tazobactam, ticarcillin/clavulanate, ceftazidime, cefepime, cefpirome aztreonam, imipenem, meropenem, ciprofloxacin and levofloxacin have been isolated. Since Acinetobacter baumannii easily become having multiple resistances and are capable of living for a while on surfaces of an object, it is a tough issue in prevention and treatment of nosocomial infections.

Phages (bacteriophages) are viruses that infect bacteria, and grow and replicate in bacteria. There are lytic phages and lysogenic phages. Lytic phages infect bacteria, replicate in bacteria, and then are released from bacteria by lysing and killing bacteria. Lysogenic phages are capable of undergoing lytic or lysogenic life cycles, and exist in host cells while in lysogenic life cycles.

It has been disclosed that bacterial diseases are treated with phages. For example, U.S. Pat. No. 5,688,501, U.S. Pat. No. 5,997,862, U.S. Pat. No. 6,248,324 and U.S. Pat. No. 6,485,902 have disclosed a pharmaceutical composition comprising phages for treating bacterial diseases, group A streptococcal infections, dermatological infections, and control of Escherichia coli O157 infections, respectively. U.S. Pat. No. 6,121,036 has disclosed a pharmaceutical composition having at least one phage. U.S. Pat. No. 6,699,701 has disclosed using Salmonella enteritidis-specific phages for packing food, in which a package material is coated with phages, and food (such as fruit and vegetables) is packed with the package material.

There are no publications disclosing Acinetobacter baumannii-specific phages, which are used for reducing the population of Acinetobacter baumannii and further reducing nosocomial infections.

SUMMARY

OF THE INVENTION

The present invention provides an isolated Acinetobacter baumannii phage, comprising one or more genomic sequences selected from the group consisting of sequences of SEQ. ID. NO: 1, 2, 3 and 4 (as shown in sequence listing), and sequences having more than 80% homology thereof.

It is known that the nucleotide sequence of RNA polymerase is a highly conserved region in viral genome, and thus homology among species can be determined by identifying homology of RNA polymerase. In the present invention, sequences of SEQ ID NO. 1 and SEQ ID NO. 2 are DNA sequences encoding RNA polymerase of Acinetobacter baumannii phages. Upon sequence alignment, there is no viral sequence in the gene bank identical or similar to the sequences of SEQ ID NO. 1 and SEQ ID NO. 2 in the present invention.

The Acinetobacter baumannii phages of the present invention were deposited in DSMZ (German Collection of Microorganisms and Cell Cultures, German), and have deposition numbers as DSM 23599 and DSM23600. In one embodiment of the present invention, Acinetobacter baumannii phages are variants of the above-mentioned deposited phages, and have genomic sequences with homology more than 80% of those in above-mentioned deposited phages.

The Acinetobacter baumannii phages of the present invention are lytic phages and specifically infect Acinetobacter baumannii. In other words, after the pages of the present invention infect host cells, Acinetobacter baumannii, the phages replicate and propagate in the host cells and lyse cell walls of host cells, and then Acinetobacter baumannii are destructed along with the release of the phages. Accordingly, the phages of the present invention are capable of reducing the amount of Acinetobacter baumannii and disinfecting environments, especially reducing nosocomial infections caused by Acinetobacter baumannii.

In an aspect of the present invention, the Acinetobacter baumannii phages are capable of attaching rapidly to Acinetobacter baumannii, have short latent period, and have large burst size upon lysis of Acinetobacter baumannii.

The phages of the present invention have double-stranded DNA having 35 to 40 kb as genetic material. FIG. 1 shows the viral particles of the phage of the present invention, in which the viral particle has a head portion with 20 faces and a tail portion have filament structure for attaching to the surface of host cells. The head portion of the viral particle is about 60 nm, and the tail portion is about 9 to 11 nm

In an aspect of the present invention, the Acinetobacter baumannii phages have acid tolerance and alkali tolerance, and have bioactivity in the environment at pH 4 to 12. In the present invention, the term “bioactivity” refers to that the pages are capable of infecting host cells, Acinetobacter baumannii, propagating in the host cells and/or lysing the host cells.

In an aspect of the present invention, the phages of the present invention have bioactivity in a surfactant.

In one embodiment of the present invention, the surfactant is one selected from the group consisting of an anionic surfactant, a cationic surfactant, an amphoteric surfactant and a non-ionic surfactant.

In one embodiment of the present invention, the anionic surfactant can be, but not limited to, ammonium dodecyl sulfate, disodium laureth sulfosuccinate, disodium octyl sulfosuccinate, linear dodecyl benzene sulfonates, dodecyl phosphates (mono alkyl phosphate, MAP), secondary alkane sulfates (SAS), sodium cocoyl isethionate (SCID), sodium lauryl ether sulfate (SLES), sodium lauroyl sarcosinate, sodium lauryl sulfate (SLS), sodium taurine cocoyl methyltaurate and so on.

In a preferred embodiment of the present invention, the cationic surfactant can be, but not limited to, cetyl trimethyl ammonium chloride, dicocodimonium chloride, didoctyl dimethyl ammonium chloride, diester quaternary ammonium salts, alkyl dimethyl benzyl ammonium chloride, ditallow dimethyl ammonium chloride (DTDMAC), imidazoline quaternary ammonium salts and so on.

In a preferred embodiment of the present invention, the amphoteric surfactant can be, but not limited to, cocoyl lmidazolinium betaine, cocoamidopropyl hydroxysultaine, cocpamidopropyl dimethyl betaine, disodium cocoamphodipropionate, lauramidopropyl betaine, sodium alkylamphopropionate, tallow dihydroxyethyl betaine and so on.

In a preferred embodiment of the present invention, the non-ionic surfactant can be, but not limited to, alkyl polygluoside (APG), cocoamide DEA, lauramine oxide, lauryl ether carboxylic acid, Triton X (such as TX-100, TX-405, etc.), PEG-150 di-stearate, Tween (such as Tween-40, Tween-80, etc.) and Span (such as Span-20, Span-80, etc.) and so on.

In a preferred embodiment of the present invention, the surfactant is a non-ionic surfactant.

In a preferred embodiment, the surfactant is a commercial product, especially a detergent.



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stats Patent Info
Application #
US 20120276612 A1
Publish Date
11/01/2012
Document #
File Date
04/17/2014
USPTO Class
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Acinetobacter


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