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Bactericide composition comprising phage

A bacteriophage and composition technology, applied in the field of bactericidal compositions, can solve problems such as no method for bacterial prevention or treatment of nosocomial infection, undisclosed bacteriophage of Acinetobacter baumannii, undisclosed Acinetobacter baumannii and the like, Achieve good effect, solve nosocomial infection, no toxic residue effect

Inactive Publication Date: 2012-02-08
HUALIEN TZU CHI HOSPITAL BUDDHIST TZU CHI MEDICAL FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] To sum up, currently known applications of phages have not yet provided effective prevention or treatment methods for nosocomial infection bacteria. Method for the number of Acinetobacter baumannii infection bacteria

Method used

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  • Bactericide composition comprising phage
  • Bactericide composition comprising phage
  • Bactericide composition comprising phage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Preparation of bacteriophage: samples collected from Hualien Tzu Chi Hospital, such as catheter washing fluid, drainage system wastewater, and untreated sewage, were centrifuged at 5,000×g for 10 minutes at 4°C, and the supernatant was 0.45-μm pore size membrane filtration followed by plaque testing.

[0052] Drop 10 μl of the sample filtrate onto the bacterial lawns of AB bacteria (in 0.7% LB agar medium) covered on the general LB agar medium. If the sample filtrate contains phage, it will be on the bacterial lawns Create a clear zone, pick it out and soak it in LB medium, filter to remove AB bacteria, and you can get a high concentration of phage. After diluting the phage, spread it on the culture medium to form a lysed plaque. Perform at least two single plaque isolation steps to obtain pure phage strains.

[0053] A total of 10 AB bacteriophage strains, ψAB1-9 and ψAB11, were isolated from the samples after identification. Among them, ψAB 1-7 and ψAB 9 have icosa...

Embodiment 2

[0055] In order to test the host specificity of the bactericidal composition of the present invention, select 127 strains of Acinetobacter baumannii (Acinetobacter baumannii, sometimes referred to as AB bacteria in this manual), wherein 125 strains are collected from Hualien Tzu Chi Hospital, and 2 strains are from ATCC (American Type Culture Collection) are ATCC 19606 and ATCC 17978 respectively.

[0056] The bacteria were cultured in LB medium (Difco Laboratories, Detroit, MI, USA) at 37°C, and the bacterial growth was monitored according to the turbidity, at 600nm (OD 600 ) to measure the absorbance value, when the OD is 1, it means that the bacterial concentration is 3×10 8 cells / ml. And a layer of 0.7% LB agar medium containing host bacteria was covered on the general LB agar medium to prepare host bacterial laws (bacterial laws).

[0057] 10 μl of phage culture solution (phage concentration of 10 10 PFU / ml) was dropped onto the bacterial layer, and the culture medium ...

Embodiment 3

[0064] When the concentration of host bacteria reaches OD 600 When the concentration is 0.6U, add AB bacteriophage (pure strain or mixture) to the host bacterial culture medium at an MOI (Multiplicity Of Infection) value of 0.0005, and cultivate at room temperature. Take 100 μl samples at the time points of 0, 1, 2, 3, 4, 5, 10, 20, and 30 minutes of incubation, dilute with 0.9 mL of cold LB, centrifuge at 12,000×g for 5 minutes, take the supernatant, and measure the The amount of phage adsorbed to the host bacteria. The result is shown in Figure 2.

[0065] Observing the host bacterial culture fluid added with phage, it was found that the culture fluid turned from cloudy to clear within 100 minutes, showing that the phage completely lysed the host bacteria, confirming that the bactericidal composition of the present invention can indeed achieve a bactericidal effect.

[0066] Depend on figure 2 It can be seen that about 75% of the phage particles adsorbed to the host bact...

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Abstract

The invention provides a bactericide composition comprising a phage, which comprises the phage and a carrier of Acinetobacter baumannii. The invention also provides a method for decontamination of medical institutions or medical-related research institutions, which is characterized in that Acinetobacter baumannii with an effective amount is applied to the medical institutions or medical-related research institutions to reduce the amount of Acinetobacter baumannii existed in the medical institutions or medical-related research institutions.

Description

technical field [0001] The invention relates to a bactericidal composition and a bactericidal method using the bactericidal composition, in particular to a bactericidal composition comprising bacteriophage and a carrier and a bactericidal method using the bactericidal composition. Background technique [0002] Nosocomial infection is the most difficult problem in hospitals. According to statistics, the nosocomial infection rate in hospitals is generally about 3% to 5%. Bacteria in nosocomial infections are usually opportunistic pathogens, that is, for hosts with normal immunity, the bacteria are harmless, and some bacteria are even normal flora on the surface of the human body; However, when the host's immunity is weakened, the bacteria can easily cause infection and disease. [0003] Bacteria that commonly cause nosocomial infections include, for example, Staphylococcus, Pseudomonas, Acinetobacter, Enterococci, Enterobacteriaceae, nonfermenting gram-negative bacilli ( Non...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/00A01P1/00A61K35/76A61P31/04A61K35/74
CPCY02A50/30
Inventor 陈立光林念璁
Owner HUALIEN TZU CHI HOSPITAL BUDDHIST TZU CHI MEDICAL FOUND
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