In the recent years, multi-drug resistance is common among many pathogenic microorganisms. This fact causes severe problems on the treatment of patients infected by such muitiple-drug resistant pathogenic microorganisms. However, the development of new effective antibiotics is very time-consuming, and last of all, a drug-resistance of such multiple-drug resistant pathogenic microorganisms against a new effective antibiotic should be expected with high likelihood.

Moreover, the scientists agree on the fact that, on the long run, the competition between the development of new effective antibiotics and the generation of resistance of pathogenic microorganisms against said new effective antibiotics will be in favour of the pathogenic microorganisms.

Thus, the technical problem underlying the present invention is to provide a new system for the treatment of diseases caused by pathogenic organisms and for the diagnosis of said diseases and the detection of said pathogenic organisms.

The above technical problem is achieved by providing a pharmaceutical composi- tion comprising a self-replicating agent directed to a heterologeneous pathogenic organism in a host, and optionally a pharmaceutically acceptable carrier and/or diluent, wherein the self-replicating agent is capable of growth-inhibiting and/or destroying the heterologeneous pathogenic organism without substantial toxicity to the host.

Further, there is provided a diagnostic kit comprising a self-replicating agent directed to a heterologenous pathogenic organism, wherein the self-replicating agent contains at least one detectable marker gene.

The aim of the invention is the infection of the respective pathogenic organism by the self-replicating agent, eventually leading to the destruction or inhibition of the pathogenic organism. This can be achieved by e.g. virulent as weil as temperate viruses. Lytic strains of viruses would be of advantage with respect to a spread through the pathogenic population which would enabie the use of low titer pharmaceutical compositions which would potentate themself in case of infectability of the pathogenic cells. The initial infection of the pathogenic host organism might be done by use of naked DNA, RNA (e.g. virions), especial- ly encapsulated nucleic acids such as viruses (e.g. eucaryotic viruses and bacteriophages), or specific systems carrying said self replicating agent. Such specific systems may be microsomes and liposomes or invasive bacteria which are already infected with said self replicating agent. Such preinfected invasive bacteria can be used for combatting infections caused by invasive pathogens such as Listeria.

A further aim of the invention is the infection of a pathogenic organism by the self-replicating agent, whereby the pathogenic organism can be detected via the marker gene contained in the self-replicating agent upon infection. The term "marker gene" means any nucleotide sequence which is introduced into the genetic material of the self-replicating agent using recombinant DNA techniques, and which can be detected upon infection of the pathogenic organism using known techniques such as PCR, enzymatic methods such as measuring the luminescence, fluorescence and/or immunologic methods. Examples of marker genes are the gene encoding luciferase, chloramphenicolacetyltransferase (CAT) and green fluorescent protein (GFP).

The term "self-replicating agent" inciudes, agents, preferably of viral origin such as eukaryotic viruses or bacteriophages, directed specifically to the heterologe- neous pathogenic organism. The self-replicating agent may be of natural origin such as naturally occurring eukaryotic viruses or bacteriophages, or may contain at least one nucleotide sequence obtained by recombinant DNA technique, such as a marker gene. Examples of such self-replicating infectious agents are na- turally occurring viruses, viroids or bacteriophages such as Listeria phages A511, B054, or Klebsieila phages K11 and Klebsiella pneumoniae bacteriophage or Staphylococcus phage group II as well as viruses infecting eukaryotic cells such as yeast L-A virus, ScVL1, ScVLa, K1 and K2 killervirus, or genetically modified versions thereof. In a preferred embodiment, the self-replication of the self-replicating agent is supported by the heterologeneous pathogenic organism.

Further examples of the self-replicating agent are Chlamydia phages such as Chp 1 (Storey etna., Gen. Virol. 70, 1989, 1321-1327; Bacon petal., Vet. Rec. 119, 1986, 618-620), Bacillus phages such as phi 20 from the Strain Sterne 34 F7 (Inal and Karunakaran, Curr. Microbiol. 32, 1996, 171-175), CP54 (Mol. Mikro- biol., Virosol. 137, 1989, 14-19) and Tgl3ant (Mol. Gen. Mikrobiol. 55, 1991, 28-31), Salmonella phages type 4 and type 8 (Zentralbl. Bakteriol. 285, 1997, 379-388) and phages 13a, 14b, 23, 8 and 2 (Singer et al., Avian, Dis. 36, 1992, 324-333) and Vibrio cholerae phage 0139. Further examples of the self- replicating agent against eukaryotic organisms are Trichonomas-RNA virus (Wang and Wang, PNAS 83, 1986, 7956-7960), Gardiavirus (GLV) (Wang et al., PNAS 90, 1 993, 8595-8599), Leishmania-dsRNA virus LRVt and LAV2.1 (Widmer and Dooley, Nucleic Acids Res. 23, 1995, 2300-2304), and Cryptosporidium- dsRNA virus (Khramtsov et al., Mol. Microbiol. 26, 1997, 289-300).

The term "heterologeneous pathogenic organism" means any organism, i.e. an eukaryotic or prokaryotic organism, which is capable of infecting a host, and thereby causing a disorder in the host during the course of infection. Such heterologeneous pathogenic organisms inciude Listeriae, Klebsiellae, Streptococ- ci, Staphylococci, Pneumococci, Moraxellae, Legionellae, Vibrio, Edwardsiellae, Yersiniae, Escherichiae, Haemophilus influenza, Proteus, Heliobacter, Salmonael- la, Chlamydia, Aeromonas salmonicida, Renibacterium salmoninarum, Candida albicans, Leishmania, Trichomonas vaginalis, Giardia lamblia, Cryptosporidium parvum, Plasmodium falciparum, Entamoeba histolytica, Pneumocystis carinii, Tryanosama rhodesiense/gambiense, Trypanosoma cruzi, and bacteriological weapons such as Bacillus anthracis. Examples are bacteria such as Listeria monocytogenesor, Klebsieila (K.) ozaenae, K. rhinoscleromatis scleromatis and K. pneumoniae, Staphylococcus aureus, Yersinia ruckeri, Renibacterium salmoni- naram, Vibrio anguillarum, Vibrio cholerae, Vibrio salmonicida, Vibrio viscosus, Edwardsiella ictalari, or fungi such as the yeast Candida albicans, or protozoa such as Leishmania. Further examples of heterologenous pathogenic organisms are Chlamydia such as Chlamydia pneumoniae, Chlamydia trachomatis and Chlamydia psittaci, Moraxella catarrhalis, Salmonella enteritides.

The term "host" includes vertebrates such as birds, fishes and mammals, prefe- rably humans.

The carrier and/or diluent may be in solid or liquid form, and do not exhibit any specific restriction, but should be stabilizing and/or protecting the self-replicating agent, and should have substantially no toxicity for the host.

The pharmaceutical composition according to the present invention may be used for the treatment of e.g. bacterial, fungal or protozoal infections of the lung such as pneumonia, infections of skin such as eczema, acne, furunkel-boils, nasal or throat infections, eye infections, infections of internal organs, ear infections or wound infections, in e.g. vertebrates such as mammals preferably in man.

Furthermore, the invention may be used for the treatment of circulating microor- ganisms in the circulatory systems such as the blood stream or lymphatic circulation, by use of e.g. infusions. Additionally, the composition according to the present invention can be used for disinfection of e.g. medicine devices such as surgical instruments, dialysis or urinary catheter or conduits, and infusion cannula or conduits. The pharmaceutical composition according to the present invention can be applied to a patient without any specific restricton regarding the amount of the self-replicating agent due to its substantial non-toxicity for the host. In a preferred embodiment of the present application the pharmaceutical composition contains a dosage of 103-109 phages/application.

The formulation of the pharmaceutical composition according to the present invention does not exhibit any specific restriction, and may be prepared e.g. in the form of tablets, suppositories, ointments, solutions, or retarded release- formulations. The pharmaceutical composition according to the present invention can be applied e.g. by way of inhalation, topically, orally, systemically, depen- dent on the formulation, to a patient suffering from an infection disease caused by a heterologeneous pathogenic microorganism.

The formulation of the pharmaceutical composition can be achieved mixing the self-replicating agents in liquid or solid form with suitable carriers. The final mixture can be applied in form of a topically cream, inhalation of a powder or mist, application of suppositories, ointments, infusion solutions, ear drops or eye drops or nose drops or sprays, or using a tape containing said mixture for topical applications.

A preferred embodiment of the present invention is a formulation as a phage- lyophilisate to be used in a powder inhalator such as the inhalator disclosed in US-patent 5,673,685. Such phage-lyophilisates may be used for the treatment of diseases caused by Klebsiella, Pneumococcus, Staphylococcus, Legionella, Haemophilus influenza, and Moraxella catarrhalis. In a further preferred embodi- ment the pharmaceutical composition of the present invention is formulated for topical applications using a substrate such as a bandage (e.g. Tegasorb, 3M) or a tape. Such bandages or tapes may be used for the treatment of diseases caused by Klebsiella, Staphylococcus, Streptococcus and Listeria. In yet another preferred embodiment the pharmaceutical composition of the present invention is formulated for oral application using e.g. gelatine capsules in gastric juice- resistant or non-resistant form. Such gelatine capsules may be used for the treatment of diseases caused by Chlamydia, Staphylococcus, Streptococcus, urinary tract infectious agents such as E. coli, Staphylococcus saprophyticus, Proteus mirabiles, Klebsiella and Heliobacter pylori.

The self-replicating agent, preferably the recombinant phage contained in the diagnostic kit according to the present invention can be used for detecting a pathogenic organism in samples derived from living organisms such as a smear, tissue, blood, urine, stool or sputum from a patient, or in samples derived from e.g. soil, air, water or other areas such as fish ponds for fish farming and/or poultry coops for e.g. chicken farming. In a preferred embodiment of the present invention the pathogenic organism contained in the sample will be infected with the recombinant phage having a suitable marker gene such as the gene encoding luciferase, chloramphenicolacetyltransferase (CAT) and green flourescent protein (GFP) upon infection the marker gene will be expressed and can be detected with known methods such as measuring the luminescence generated by the expressed luciferase.

Examples of pathogenic organisms to be detected are as listed above. Preferred examples of pathogenic organisms are Chlamydia such as Chlamydia pneumo- niae, Chlamydiatrachomatis and Chlamydia psittaci, Pneumococci, Haemophilus influenza, Staphylococci such as St. aureus, Moraxellae such as M. catarrhalis, Legionellae, Streptococci, Salmonellae such as Salmonella enteritides, Aeromo- nas salmonicida, and bacteriological weapons such as Bacillus anthracis.

For diagnostic use the self-replicating agent can be admixed with any suitable carrier and/or diluent in solid or liquid form, but preferably should stabilize and/or protect the self-replicating agent.

A further subject of the present invention is a process for the preparation of the above mentioned pharmaceutical composition, comprising the steps of: (a) sampling the heterologeneous pathogenic organism from a specimen of the host, (b) inoculating the heterologeneous pathogenic organism with one or more types of self-replicating agents, (c) selecting and/or adapting the desired self-replicating agent having a high growth-inhibiting and/or destroying effect, and (d) formulating the pharmaceutical composition.