TW201105241A - Bactericide composition comprising a phage - Google Patents

Abstract

This invention provides a composition for killing bacteria in vitro comprising a phage for infecting Acinetobacter baumannii and a carrier. This invention also provides a method for decontamination of medical institution or medical-related research institution, which comprises applying effective amount of phage for infecting Acinetobacter baumannii to the medical institution or medical-related research institution to reduce amount of Acinetobacter baumannii existed in the medical institution or medical-related research institution.

Description

201105241 - VI. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a bactericidal composition and a sterilizing method using the sterilizing composition, in particular to a bactericidal composition comprising a bacteriophage and a carrier, and an application thereof The sterilization method of the sterilization composition. [Prior Art] Nosocomial infection is the most difficult problem in hospitals. According to statistics, the hospital's in-hospital infection rate is about +3% to 5%. In-hospital • Infected bacteria are usually 饲pp0rtunistic pathogens, that is, for normal immune hosts, these bacteria are harmless, and some bacteria are even normal flora on the surface of the human body. However, when the host's immunity declines, the bacteria are prone to cause infection and cause disease. Common nosocomial infections include, for example, Staphylococcus, Pseudomonas (ΡπΜί/omo(10)?), Acinetobacter φ (Jc/zzeioiflcier), Enterococcus, Enterobacteriaceae, Non-fermenting Gram-negative bacilli (Non -fermentative gram-negative bacilli), Legionella genus, Clostridium (C/c^irWz'wm), Mycobacterium {Mycobacterium) # 〇 Currently, the most common nosocomial infections include Pseudomonas aeruginosa ), Staphylococcus aureus (flwrews) and Acinetobacter baumannii (baumannii), etc. 0 3 111057 201105241 Nosocomial infections may survive in stethoscopes, medical records, tourniquets, gloves, needles, respirators, moist bottles, furniture, flooring, ventilation Equipment such as mouth, monitor, etc.; or in water, soil and food (fruits, vegetables) and sewers; or in the human body, such as the skin, underarms, conjunctiva, mouth, upper respiratory tract, nasopharynx and gastrointestinal tract Wait. The incidence of nosocomial infections, in the case of intensive care units, because the patients are mostly critically ill patients, the human body immunity is poor, and often need to accept invasive treatment, such as intubation, vascular devices, etc., greatly improve the possibility of nosocomial infection, according to statistics, The infection rate in the intensive care unit is about 20 to 30 per 1,000. The treatment of common bacterial infections is the use of antibiotics. However, due to the abuse of antibiotics, bacteria are screened to evolve more resistance. Currently, there are more and more bacteria resistant to antibiotics in nosocomial infections. To treat patients infected with this bacterium, expensive new antibiotics must be used, and if the resistance of the bacteria continues to develop, no effective antibiotics will be available. In view of this, it is necessary to develop alternative methods for reducing and/or preventing nosocomial infections. The phage (bacteriophage) is a kind of virus characterized in that the host of the phage is a bacterium and must be grown and replicated in the bacterium. The bacillus can be divided into a lytic type and a lysogenic type. The lytic phage will infect the host bacteria. After the replication in the host is completed, the phage will lyse and release the bacteria, and the bacteria will rupture and die. Protoplasts are milder phage that can undergo a lytic or solvate life history and coexist with the host in the path of the protoplast. The use of bacteriophage for the treatment of bacterial diseases has been previously described, for example, in U.S. Patent Nos. 5,688,501, 5,997,862, et al., 6,248,324, 6, 485, 902, respectively, to disclose the use of medicinal compositions comprising bacteriophage to treat bacteria. Sexual diseases, Streptococcus type A (5^印 (3) aw A), bacteria causing skin infection, Escherichia coli 〇157 strain, etc. U.S. Patent No. 6,m,036 discloses a medical treatment comprising more than one type of bacillus Compositions; U.S. Patent No. 6, _, 701 discloses the use of Salmonella rods = (Mm (10) _ specific phage for food packaging methods ^ (4) sputum bacteria coated onto packaging materials, and then coated with packaging materials ( Such as sparse fruit.), above = the current known (four) body application, has not provided a method for the prevention or treatment of the hospital's feelings, Tian Hao, effective prevention, treatment, no disclosure of Acinetobacter baumannii, then salty The bacteria (4)^ also did not disclose the method for reducing the number of Acinetobacter baumannii in the hospital. [Invention content] ° To overcome the above and other problems, the composition of the bacteria , including to + one #毛月仏犍 for an in vitro killing such as dirty _) 嗟 体: Acinetobacter baumannii (agent, wherein the sputum bacteria in the two 'the sputum system contains at least two does not move: In the state of the invention, the lion, the bacterial body. In the - state, the sputum: with nuclear tolerance and tolerance to the underlying parts of the bacterium to keep the bacteria; ^ coffee above to _ In the following article, the biological activity of 誃-嗟 体 谨 谨 可 可 可 可 111 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 057 : the infectivity of the host Acinetobacter baumannii, capable of infecting the growth of the sputum, the cytoplasm, and/or the ability to lyse the host cells. Bowen: The lysate-type bacterium of the present invention is infected After the bacterial cell (4) is completed, it can be lysed to eliminate the second cell of the rod and release the proliferation, and the A. baumannii is inhibited. 9 This invention can be used to inhibit Acinetobacter baumannii/ten Or reducing the amount of Acinetobacter baumannii. In the composition of the present invention, the carrier is not specially prepared, as long as it contains you AB bacteria To be compatible -p_e) Oh enabling the cells to be retained in the carrier the biologically active agent 'was used in the sterilizing composition. In one aspect, the carrier is in the form of a solution, a suspension, a powder, a spray, or an ointment, but is not limited to the implementation of the carrier, the carrier is: oil, surfactant (such as cleaning) Agent, soap), protein gland (Qing (4) Temple: can also be a combination of two or more, of which water and / or surfactant is the best car. In a preferred embodiment, the surfactant is At least one surfactant selected from the group consisting of an anionic surfactant, a cationic boundary agent, an ionic surfactant, and a nonionic surfactant. In an embodiment, the anionic interface The active agent is, by way of example and not limitation, ammonium lauryl sulfate, disodium laureth sulfosuccinate, disodium octylsulfonyl succinate, soft dodecylbenzene sulfonic acid, twelve Alkyl phosphate (MAP), secondary alkyl phosphate (SAS), cocoyl hydroxyethyl]]] 〇 57 6 201105241, sodium citrate (SCID), sodium lauryl sulfate (SLES), Lauric acid muscle · sodium amide, sodium lauryl ether sulfate (SLS), sodium sulfonate In the above examples, the cationic surfactant is, by way of example and not limitation, Cetyl trimethyl ammonium oxide, dicocoyl dimethyl ammonium chloride, dimercapto dimethyl Ammonium chloride, diester quaternary ammonium salt, alkylbenzyl dimethyl emulsifier, tallow alkyl dimercapto chloride (DTDMAC), ° m beta sedentary quaternary ammonium salt, etc. In the examples , the zwitterionic surfactant is, for example, but not the same as the 'or oil-based σ m α sitting on the sweet test (c〇c〇yi imidaz〇liniuni betaine), cocoamphetamine hydroxy sulfobeet Alkali, cocoamidopropyl dimethyl betaine, cocoamphodipropionate di-nano, lauryl propyl dimethyl sweetness test, sodium alkylamphopropionate, tallow Dihydroxyethyl betaines, etc. In the examples, the nonionic surfactant is, for example, but not limited to, 'APG, agglomerate (cocoamide DEA), • Laurel Amine oxides, lauryl ether carboxylates, TritonX (eg TX-100, TX-405, etc.), PEG-150 di-stearate, Tween (eg Tween-40, T Ween-80, etc.), Span (such as Span-20, Span-80, etc.), etc. In the preferred embodiment, the surfactant may be a commercially available product, particularly a detergent product. The composition of the present invention is as needed. The second phage is included, wherein the second phage system is a phage which infects bacteria in the hospital. In one aspect, the hospital infection virus is selected from the group consisting of Acinetobacter sp. r, Staphylococcus (10), Enterococcus 7 111057 1 201105241 ( £>Uerococcz·), Enterobacteriaceae, Non-fermentative gram-negative bacilli, Legionnaires' disease (/^/(10)hex//^), Clostridium, points One or more of the group consisting of Mycobacterium (Afycc) Z?acieWwm) and Pseudomonas. In an embodiment, the Acinetobacter is selected from one or more of the group consisting of A. calcoaceticus 'A. haemolyticus ' A. junii ' i and 乂. of Acinetobacter baumannii (left baumannii). In the examples 'the staphylococcus strain is selected from iS.ep/cfermzWz's, S. hemolyticus, S. auricularis - S. capitis, S. caprae, S. hominis, S. pasteuri ' S. simulans, S. warneri, S One or more of the groups consisting of .cohnii, S.sciuri, and *. In an embodiment, the Enterobacteriaceae is selected from the group consisting of Escherichia coli (£^c/zerzc/n<2), Klebsiella pneumoniae (), Proteus, Enterobacter (jEViiaohacie/", Citrobacter ( One or more of the group consisting of Morgime//fl, Salmonella, Serratia (), Shigella (), and Yersinia. In the embodiment, the Pseudomonas is Pseudomonas aeruginosa. The second phage system contained in the composition of the present invention is a lytic phage which infects a host (bacteria) and is in a bacterial cell. After the completion of the proliferation, the phage which releases the proliferating phage can be lysed by the cell wall of the host, and the host dies, whereby the second phage of the present invention can be used to inhibit the activity of the host or reduce the number of the host. In one aspect, the second phage retains the biological activity of the phage in the surfactant. In one aspect, the initial concentration of the Acinetobacter baumannii phage in the composition is lxl〇7PFU/ml to lxl09PFU/m, thereby allowing the bacteriophage of the AB phage and other bacteria according to the needs of the user. The composition of the present invention is used to prepare the composition of the present invention, whereby the effect of simultaneously reducing a plurality of bacteria can be attained. In another aspect, the present invention provides an in vitro sterilization method for a medical institution or a medical related research institution, comprising: applying a bactericidal composition comprising a bacillus of Acinetobacter baumannii and a carrier; The medical institution or medical research institution to reduce the number of bacteria in the medical institution or medical research institution. In one aspect, the φ method of the present invention is particularly directed to reducing the amount of Acinetobacter baumannii. Wherein the phage of Acinetobacter baumannii and the carrier are as defined above. The bactericidal composition further comprises a second phage as defined above, as desired. Thereby, the composition of the present invention can be prepared by the combination of AB bacteria phage alone or with other bacterial phage according to the needs of the user, thereby reducing the number of Acinetobacter baumannii or simultaneously reducing the number of various bacteria. The effect. In one aspect, the bactericidal composition is applied to a medical facility or a medical-related research institution, such as a hospital or a nursing home. In an embodiment

9 111057 L 201105241, the bactericidal composition is applied to the environment of the hospital or nursing home, for example, but not limited to, an intensive care unit, an operating room, a recovery room, a treatment room, a reception room, etc.; or in the hospital or The equipment of the nursing home, for example, but not limited to, the surface of the cannula, vascular device, stethoscope, medical record, tourniquet, gloves, respirator, moist bottle, furniture, floor, vent, monitor, and the like. The method of administration is selected depending on the subject matter to be administered, by direct spraying, indirect spraying, soaking or smearing. The bactericidal composition and the sterilizing method of the invention can effectively reduce the number of the target bacteria or inhibit the activity of the target bacteria by utilizing the specificity between the phage and the host, thereby solving the problem that the antibiotic resistance can not be used due to the antibiotic resistance of the bacteria. Moreover, the use of the bactericidal composition of the present invention does not cause further resistance of the target bacteria. The bactericidal composition of the present invention can be used in combination with a commercially available detergent, and has an advantage in commercialization. In addition, the sterilization method of the present invention has simple steps and good effects, and has no doubt that the chemical germicide is toxic and has no bacterial resistance, so that the problem of nosocomial infection can be effectively solved. [Embodiment] The embodiments of the present invention are described by way of specific examples, and those skilled in the art can understand the other advantages and effects of the present invention from the disclosure of the present disclosure. Example 1 Preparation of phage: Pipette washing liquid collected from Hualien Tzu Chi Hospital, wastewater of untreated sewage, etc. Samples were centrifuged at 5,000 x g for 10 minutes at 4 ° C, and the supernatant was centrifuged. The plaque test was followed by a 0.45-/zm pore size of 10 111057 201105241 ·- Membrane filtration ''. - Drop the sample of 10 #1 to the bacterial iawns of AB bacteria (used in 7% LB agar medium) on the general LB agar medium. If the sample filtrate contains bacteriophage, it will A clear zone is created on the bacterial layer. 'Pick it out and immerse it in lb medium, and filter to remove AB bacteria to obtain a high concentration of sputum cells. The sputum was diluted and applied to the medium to form plaque. A single plaque detachment step is performed at least twice to obtain a phage pure strain.

A total of 5 AB steroids were obtained after identification of the AB phage phage isolated from the sample, and each strain was infected with AB bacteria, and the infectivity of each strain of AB strain was slightly different. Example 2 In order to test the host specificity of the bactericidal composition of the present invention, the scorpion species as shown in Table 1 was selected, wherein 'Acinetobacter baumannii (6(10) mawm'z·, sometimes referred to as AB bacteria in the present specification). Source: 35 strains Collected from the Chunhualian Tzu Chi Hospital's 2 strains from the ATCC (American Type Culture Collection), cultured the bacteria in LB medium (Difco Laboratories, Detroit, MI, USA) at 37 ° C, and monitored the growth of bacteria by turbidity. The absorbance was measured at 600 nm (OD600), and the OD was 1. The bacterial concentration was 3 x 1008 cells/ml. The host LB agar medium was covered with a layer of 0.7% LB agar medium containing host bacteria (e.g., the strain in Table 1), thereby preparing host bacterial lawns. The 10# 1 phage (pure strain or mixture) culture solution (phage rich 11 111057 1 201105241 degrees 101 GPFU / ml) was dropped into the bacterial layer, and the culture plate was dried in a sterile laminar flow table for 10 minutes at 37 ° C. Incubate for 18 to 20 hours and observe whether plaque is produced or not. Table 1 Bacterial name Characteristics _ Source Strain _

Acinetobacter baumannii

19606,17987 ATCC Standard strain ATCC Hualien Tzu Chi Hospital M495, Ml094, M2472, clinical strain, MDRAB M2477, M2641, M2835, M3069, M3237, M3739, M3982, M4666, M5473, M67329, M67649, M67777, M68092, M68282, M68316 , M68630, M68651, M68653, M68661, 45530, 46709, 47538, 47543, 48393, 48465, 48829, 49575, 50064, 50068, 50913, 51360 M2383 Clinical strain

Abl-Ab9

Imir Merr Ampr Hualien Tzu Chi Hospital Wu, et al. (2007)

Acinetobacter cal coaceticus

ATCC 33305 ATCC Standard Strain 12 111057 201105241

Escherichia coli DH5a endAl hsdRl 7 (rk-wk-l·) supE44 thi-1 recAl gyrA Γθ1Α1ψ80ά lacZAM15A (lacZYA-argF)U169 Hanahan D. (1983) G0003, G0004, G0008, G0010, G0012, G0070, GOOTi, G0072, G0081 Clinical strain Klebsiella pneumoniae Kp2, Kp50, Kp53, Kp90? Kpl20, Kpl21 Clinical strain Wu, et al. (2007) Pseudomonas aeruginosa Pa79, Pa81, Pa86 Clinical strain Wu, et al. (2007) MDRAB: Multidrug resistance AB bacteria, for gentamicin, amikacin, piperaci 11 in/tazobactam, ticarcillin/clavulanate, ceftazidime, cefepime Cefepime), cefpirome, aztreonam, imipenem, meropenem, ciprofloxacin, and levofloxacin are resistant.

Amp: ampicillin; Imi: imipenem; Mer: meropenem; r: drug resistance; s: sensitivity. The results showed that the AB bacteriophage (pure strain or scorpion conjugate) prepared according to the method of Example 1 was used in 10. strain cfl/coacei/cws, _£"· co/ζ·, strain, .6 The plaques were not produced on the bacterial layer of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains of the strains The bacteria have host specificity. 13 111057 201105241 In addition, the AB bacillus (pure strain or mixture) prepared according to the method of Example 1 has the infectivity in addition to the two standard strains of ATCC, and is also provided for the clinically isolated multi-drug resistant strain AB. Infectivity can indeed solve the problem of nosocomial infection and multi-drug resistant AB infection. Example 3 When the host bacterial concentration reached 〇D600 of 0.6 U, the AB bacteriophage (pure strain or mixture) was added to the host bacterial culture solution at a MOI (Multiplicity Of Infection) value of 0.0005, and cultured at room temperature. Sample 100/z 1 at 0, 1, 2, 3, 4, 5, 10, 20, 30 minutes of culture and dilute with 0.9 mL of cold LB, centrifuge at 12,000 xg for 5 minutes, and take the supernatant. The amount of phage that was not adsorbed to the host bacteria was determined. The result is shown in Figure 1. The host bacterial culture solution to which the phage was added was observed, and it was found that the culture solution was changed from turbidity to clarification within 100 minutes, indicating that the phage completely lysed the host bacteria, and it was confirmed that the bactericidal composition of the present invention can achieve the bactericidal effect. As can be seen from Fig. 1, about 75% of the phage particles were adsorbed to the host bacteria in 2 minutes, and about 95% of the phage particles were adsorbed to the host bacteria in 4 minutes, and reached 100% adsorption at 10 minutes. In addition, the replication curve of the bacillus was determined by a one-step growth curve, and the host bacteria of 〇D6〇o was 〇·8 U were centrifuged, and the cells were harvested, and then re-dissolved in 0.8 ml of LB medium. The concentration was 109 CFU/ml. The AB bacteriophage was added to the host bacterial culture solution at an MOI value of 0.0001, and placed at 4 ° C for 30 minutes to allow the phage to adsorb to the host bacteria. The mixture was centrifuged at 12,000 x g for 10 minutes, and the pellet containing the infected bacteria of 14 111057 201105241 ' was re-dissolved in 20 ml of LB medium, and cultured at 37 ° C. 'Sampling every 5 minutes' and the sample was immediately Dilute and fix. The result is shown in Figure 2. The incubation period is defined as self-adsorption (10 minutes without pretreatment) and the first burst (burst, release of bacteria-dissolving bacteria), as shown in Figure 2 with an incubation period of 15 minutes. The average burst was calculated to be approximately 200 PFU/cell in proportion to the ratio of the final phage particle size to the initial amount of infected bacteria. ® Therefore, the present embodiment demonstrates that the bactericidal composition of the present invention has a rapid bactericidal effect, is short in time, and has good effects; and can release a larger amount of sputum bacteria to the environment after lysing the host bacteria, so that the bactericidal effect can be continued. presence. Example 4 The invention was prepared with the surfactants TWEEN 20, TWEEN 80 and Triton X-100 (purchased from Sigma-Aldrich Biotechnology, USA) and the AB bacteriophage (pure strain or mixture) isolated from Example 1. Sterilizing composition. The conventional concentration of the conventional surfactant is mostly between 0.1 and 1 wt%. Therefore, when preparing the bactericidal composition of the present invention, 0.1 to 1 wt% of the surfactant is included, and the initial concentration is 5 x 107 PFU/ml. The phage was mixed and placed at room temperature. The phage concentration was sampled every 24 hours, and the survival fraction was calculated as follows: Phage survival rate = sampled phage concentration / phage original concentration to determine interfacial activity The effect of the agent, when the surfactant was 1 wt%, is shown in Figure 3. 15 111057 201105241 It has been determined that 0.1% to 1% by weight of surfactant is not affected by the activity of AB bacteriophage. In addition, as can be seen from Fig. 3, the AB strain is most stable in Triton X-100, TWEEN 20 times; while in TWEEN 80, although the survival rate of AB bacteria is large, it can still maintain enough. Infecting host bacteria for activity. And as time progresses, the trend of decreased phage concentration tends to moderate and gradually increase. The CV values of the three surfactants were all less than 20% as assessed by the coefficient of variation. Therefore, it was confirmed that the phage was very stable among the three surfactants. It is shown that the bactericidal composition of the present invention is diazepam. Example 5 The stability of the bactericidal composition of the present invention was determined under different environmental conditions. (1) Temperature The phage is diluted to 1〇8 PFU/ml with sterile water and placed under different temperature conditions, 4°C, 25°C, 37°C, 42°C, -20°C, and -80 ° C. At 4 ° C, 25 ° C and 37 ° C, the phage concentration was sampled every 3 hours for 24 hours, and then continued until 12 weeks per week. The results are shown in Fig. 4A. The experiments at -20 °C and -80 °C were divided into two groups. The first group was repeated freeze-thaw, the tracking time was 12 weeks, the second group was only thawed once, and the tracking time was 5 weeks. The result was 4B. The figure shows. (2) pH: The phage was diluted with an acidic (pH 4) and alkaline (pH 11) aqueous solution to a phage concentration of 1〇8 PFU/ml, and in experiments with pH values of 4.7, 7, and 11 The concentration was measured every 3 hours during the hour, and then 16 111057 201105241 was tracked once a week for 12 weeks. The results are shown in Figure 5. (3) Chemical substance The phage was added to a chloroform solution (0.5% and 2%) to a concentration of 108 PFU/ml, and the concentration was measured every 3 hours for 24 hours, and then fixed in a 0.5% chloroform solution per week. Track once, continuously for 3 weeks; 2% of the chloroform solution was followed for 6 weeks, and the results are shown in Figure 6. (4) Drying treatment The 1010 PFU/ml phage were divided into two groups, A and B groups. Group A was treated with peptone. Group B was diluted 10 times with sterile water to separate the sputum concentration by vacuum centrifugal drying system. (speed vac) drying treatment, the dried A and B groups were re-dissolved in 0.5 ml of peptone and 0.5 ml of sterile water respectively, and the concentration changes of the phage before and after drying were observed. The results are shown in Table 2. Table 2 After phage drying, the total concentration of phage after concentrated reconstitution (PFU/ml) degree (PFU/ml) body viability A group 2.18xl09 1.02χ1010 21.3% Β group 2.30χ109 1.02xl010 33.4% by the above test As a result, it was found that the bacteriophage of the present invention can survive for at least 8 weeks at a low temperature (-20 ° C, -80 ° C, 4 ° C) and the survival rate is over 5%. At ambient temperature (25 ° C and 37 ° C), the phage can survive for more than 11 weeks and the survival rate is above 14.9%. While tracking for 2 weeks at a high temperature of 17 111057 201105241 (42 ° C), the phage survival rate still reached 14.8%. In the alkaline (pH=11) environment of the bactericidal composition of the present invention, the survival rate of the phage can be maintained by about 30% after about 11 weeks, while in the acidic (pH=4) environment, the survival can still be measured until the 11th week. Phage. In addition, the AB bacteriophage survival rate measured after vacuum drying and reconstitution was as high as 20% or more. Further, in the bactericidal composition of the present invention, in 0.5% and 2% of the simulated solution, the phage can survive for more than 3 weeks and the survival rate is 30%. In summary, the phage can maintain the survival rate for the ambient temperature, dry humidity, pH value and chemical substance, and the stability of the bactericidal composition of the present invention is confirmed. Example 6 Sterilization test conditions: bacteria of the same concentration (107 CFU/ml) were placed in a container of the same specification, and the bactericidal composition of the present invention comprising 107 PFU/ml of phage was sprayed into a container, and samples were taken at different time points. The bacterial and phage concentrations were separately measured according to standard procedures, and the bactericidal effect was determined by the amount of bacterial reduction. The components of the bactericidal composition of the present invention may include the following components: (1) AB bacteriophage and sterile water (2) AB bacteriophage and surfactant (3) AB bacillus, sterile water, and surfactant (4) AB bacillus, second phage (staphylococcus), and sterile water (5) AB bacillus, second phage (Staphylococcus 18 Π1057 201105241, () sputum) And surfactants (6) AB bacteriophage, second phage (staphylococcus (bacteria), sterile water, and surfactants. In addition, spray sterile water as a control group. It was observed that the mixture in the container would be turbid. Turning to clarification, it is shown that the phage completely lyse the host bacteria, and can effectively reduce the amount of bacteria up to several logs, confirming that the bactericidal composition of the present invention can achieve the bactericidal effect. * The above examples are merely illustrative. The composition and the preparation method of the present invention are not intended to limit the present invention, and those skilled in the art can modify and modify the above embodiments without departing from the spirit and scope of the present invention. The scope of protection of the invention is as set forth in the scope of the patent application described below. [Simplified description of the drawings] Fig. 1 is the adsorption rate of Acinetobacter baumannii phage to host bacteria, and Fig. 2 is the Acinetobacter baumannii bacillus One-step growth curve; Figure 3 is the activity of Acinetobacter baumannii phage in the surfactant; Figure 4A shows the stability of the bactericidal composition of the present invention under different temperature conditions; Figure 4B shows the difference The stability of the bactericidal composition of the present invention under temperature and thawing conditions; Figure 5 illustrates the stability of the bactericidal composition 19 Π 1057 201105241 of the present invention under different pH conditions; and Figure 6 illustrates the presence of a chemical substance in the presence of a chemical substance. The stability of the bactericidal composition of the invention [Description of main component symbols] 〇20 111057

Claims (1)

201105241 VII. Scope of application for patents: 1. An in vitro bactericidal composition, comprising: Campylobacter sinensis (ribs//) sputum bacteria and sputum, carrier; wherein the phage is compatible with the carrier Sexually, to maintain the biological activity of Acinetobacter baumannii phage. 7 ^ 2·=Please patent scope! The composition of the item, wherein the Qianshi 3 is dry and the scorpion body contains at least one pure Acinetobacter baumannii bacillus. • Please refer to the composition of item 1 of the patent scope, in which the Bauer does not move to the bacteria. The witchworm system is specifically infected with Acinetobacter baumannii. 4 t Please consult the composition of the first item of the patent scope, in which the rhyme does not move, and the bacteriophage phage system is a lytic phage. \If the composition of the third item of the phase range, the towel, the Acinetobacter baumannii bacterium retains the biological activity of the bacillus in the condition of pH 4 to U. )·If you apply for a patent scope! The composition of the item, wherein the carrier is in the form of a solution, a night suspension, a powder, a spray or an ointment. The composition of claim 1, wherein the carrier is water, oil, a surfactant, a protein gland or a combination thereof. ;. For example, the composition of the patent (4) 7 items, among them. The surfactant is at least one surfactant selected from the group consisting of an anionic surfactant, a cationic surfactant, a zwitterionic surfactant, and a nonionic surfactant. (1) 057 201105241 9. The composition of claim 7, wherein the surfactant is a nonionic surfactant. 10. The composition according to claim 1, wherein the second phage is a bacteriophage which infects bacteria in the hospital. 11. The composition of claim 10, wherein the infective bacteria in the hospital are selected from the group consisting of acinetobacter (dcheiokcier), staphylococcus (), enterococci (hierococc/), Enterobacteriaceae (Enterobacteriaceae), non-fermented leather One of a group consisting of Non-fermentative gram-negative bacilli, Legionella, Clostridiwn, Mycobacterium genus (M less, and Pseudomonas) Or more. 12. The composition of claim 1 wherein the initial concentration of the Acinetobacter baumannii phage in the composition is from 1 x 10 〇 7 PFU/ml to 1 x 10 9 PFU/m b. An in vitro sterilization method of a medical institution or a medical research institution, comprising administering an effective amount of the bactericidal composition according to any one of claims 1 to 11 to the medical institution or a medical research institution To reduce the number of bacteria in the medical institution or the medical research institution. 14. The method of claim 13, wherein the application method is direct spray, indirect spray , Soaking or painting.]]] 〇57