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Methods and Kits for Detecting Fungal Infection Verfahren Und Kits Zum Nachweis Einer Pilzinfektion Procédés Et Trousses Pour Détecter Une Infection Fongique

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Methods and Kits for Detecting Fungal Infection Verfahren Und Kits Zum Nachweis Einer Pilzinfektion Procédés Et Trousses Pour Détecter Une Infection Fongique (19) TZZ_Z_T (11) EP 1 955 069 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 9/127 (2006.01) G01N 21/00 (2006.01) 18.03.2015 Bulletin 2015/12 G01N 33/53 (2006.01) G01N 33/569 (2006.01) (21) Application number: 06821559.9 (86) International application number: PCT/IL2006/001318 (22) Date of filing: 15.11.2006 (87) International publication number: WO 2007/057891 (24.05.2007 Gazette 2007/21) (54) METHODS AND KITS FOR DETECTING FUNGAL INFECTION VERFAHREN UND KITS ZUM NACHWEIS EINER PILZINFEKTION PROCÉDÉS ET TROUSSES POUR DÉTECTER UNE INFECTION FONGIQUE (84) Designated Contracting States: • TEJADA-SIMON M V ET AL: "Production of AT BE BG CH CY CZ DE DK EE ES FI FR GB GR polyclonal antibody against ergosterol HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI hemisuccinate using Freund’s and Titermax SK TR adjuvants" JOURNAL OF FOOD PROTECTION, DES MOINES, IO, US, vol. 61, no. 8, 1 August 1998 (30) Priority: 15.11.2005 US 736814 P (1998-08-01) , pages 1060-1063, XP009125825 ISSN: 0362-028X (43) Date of publication of application: • WALKER-CAPRIOGLIO H M ET AL: 13.08.2008 Bulletin 2008/33 "ANTIBODIES TO NYSTATIN DEMONSTRATE POLYENE STEROL SPECIFICITY AND ALLOW (73) Proprietor: Semorex Inc. IMMUNOLABELING OF STEROLS IN North Brunswick, NJ 08902 (US) SACCHAROMYCES CEREVISIAE" ANTIMICROBIAL AGENTS AND (72) Inventors: CHEMOTHERAPY, AMERICAN SOCIETY FOR • GREEN, Bernard, S. MICROBIOLOGY, WASHINGTON, DC, US, vol. 33, 76229 Rechovot (IL) no. 12, 1 December 1989 (1989-12-01), pages • TZOMIK, Inna 2092-2095, XP000999504 ISSN: 0066-4804 97299 Modiin (IL) • PALLAVI R M V ET AL: "Synthesis of the antigen • ARAD-YELLIN, Rina bovine serum albumin-ergosterol and its 76603 Rechovot (IL) immunocharacterization" FOOD AND AGRICULTURAL IMMUNOLOGY, XX, XX, vol. 9, (74) Representative: Dennemeyer & Associates S.A. no. 2, 1 January 1997 (1997-01-01) , pages 85-95, Poccistrasse 11 XP009125846 ISSN: 0954-0105 80336 München (DE) • WALKER-CAPRIOGLIO H M ET AL: "ANTIBODIES TO THE POLYENE ANTIMYCOTIC (56) References cited: NYSTATIN ALLOW SPECIFIC US-A1- 2002 018 808 US-B1- 6 224 902 IMMUNOLABELLING OF ERGOSTEROL IN US-B1- 7 087 395 SACCHAROMYCES-CEREVISIAE" ABSTRACTS OF THE ANNUAL MEETING OF THE AMERICAN SOCIETY FORMICROBIOLOGY, WASHINGTON, DC, US, vol. 89, 1 January 1989 (1989-01-01), page 256, XP009125845 ISSN: 0094-8519 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 1 955 069 B1 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 1 955 069 B1 • ROBERT E DALES ET AL: "Testing the • WALKER-CAPRIOGLIO H. ET AL.: ’Antibodies to association between residential fungus and Nystatin Demonstrate Polyene Sterol Specificity health using ergosterol measures and cough and Allow Immunolabeling of Sterols in recordings" MYCOPATHOLOGIA, KLUWER Saccharomyces cerevisiae’ ANTIMICROBIAL ACADEMIC PUBLISHERS, DO, vol. 147, no. 1, 1 AGENTS AND CHEMOTHERAPY vol. 33, no. 12, October 1999 (1999-10-01), pages 21-27, December 1989, pages 2092 - 2095, XP000999504 XP019259485 ISSN: 1573-0832 • DATABASE WPI Derwent Publications Ltd., • MATSUMORI NOBUAKI ET AL: "An amphotericin London, GB; AN 1999-562922, XP008117464 & DE B-ergosterol covalent conjugate with powerful 198 14 815 A1 (DUEZGUENES ET AL) 07 October membrane permeabilizing activity." CHEMISTRY 1997 & BIOLOGY MAY 2004, vol. 11, no. 5, May 2004 (2004-05), pages 673-679, XP002556042 ISSN: 1074-5521 2 EP 1 955 069 B1 Description [0001] The present invention relates to the use of a dienophile compound for determining a presence and/or a level of an ergosterol-containing organism in a substrate according to claim 1, a kit suitable for determining a presence and/or 5 a level of an ergosterol-containing organism in a substrate according to claim 4. [0002] The incidence of fungal infections and mycoses has increased significantly in the past two decades, mainly due to the growing number of individuals who have reduced immunological function (immuno-compromised patients), such as cancer patients, patients who have undergone organ transplantation, patients with AIDS, patients undergoing hemodialysis, critically ill patients, patients after major surgery, patients with catheters, patients suffering from severe 10 trauma or bums, patients having debilitative metabolic illnesses such as diabetes mellitus, persons whose blood is exposed to environmental microbes such as individuals having indwelling intravenous tubes, and even in some elderly individuals. Fungal infections are often also attributed to the frequent use of cytotoxic and/or antibacterial drugs, which alter the normal bacterial flora. [0003] Fungi include moulds, yeasts and higher fungi. All fungi are eukaryotic and have sterols but not peptidoglycan 15 in their cell membrane. They are chemoheterotrophs (requiring organic nutrition) and most are aerobic. Many fungi are also saprophytes (living off dead organic matter) in soil and water and acquire their food by absorption. Characteristically fungi also produce sexual and asexual spores. There are over 100,000 species recognized, with 100 infectious members for humans. [0004] Human fungal infections are uncommon in generally healthy persons, being confined to conditions such as 20 Candidiasis (thrush) and dermatophyte skin infections such as athlete’s foot. Nevertheless, yeast and other fungi infec- tions are one of the human ailments which still present a formidable challenge to modem medicine. In an immuno- compromised host, a variety of normally mild or nonpathogenic fungi can cause potentially fatal infections. Furthermore, the relative ease with which human can now travel around the world provides the means for unusual fungal infections to be imported from place to place. Therefore, wild and resistant strains of fungi are considered to be one of the most 25 threatening and frequent cause of death mainly in hospitalized persons and immuno-compromised patients. [0005] Invasive fungal infection (IFI) is a serious and potentially life threatening disease that affects a growing number of patients. The projected average incidence of systemic fungal infections in the United States is 306 per million, with Candidiasis accounting for 75 % of the reported cases [see, for example, Wilson et al. Value in Health, 5, 26-34, 2002]. [0006] Mortality rates in cancer patients who develop systemic fungal infections are very high. It has been observed 30 that fungi are the most common cause of nonbacterial infection in patients with leukemia and lymphoma, with Candida species and Aspergillus being the most common fungal species in cancer patients. These two infections are estimated to have a combined mortality of 20 % (Lopez-Berestein et al., Cancer Drug Delivery, 1:37-42, 1983). In other cases, fungal or fungus-like infections, usually introduced into the lungs through the air, are commonplace among large numbers of persons due to environmental exposures. 35 [0007] Certain other organisms that have parasitic properties, such as leishmaniasis, can mimic many of the disease- causing properties, behaviors, and pathologies of fungal infections. [0008] Accurate data regarding the incidence of systemic mycoses and associated mortality are difficult to obtain because reporting requirements vary; many fungal-related deaths are not reported as such because they are undiag- nosed, misdiagnosed, or not specified as cause of death. Nevertheless, many indications suggest that the incidence of 40 fungal infections and their attributable mortality are rising. This reflects the increasing number of susceptible hosts due to factors such as the HIV epidemic, advances in organ transplantation and cancer chemotherapy, and the increasing use of invasive procedures for treatment, monitoring, and life support. Estimates are that among the 35 million patients admitted to hospitals in the United States each year, at least 2.5 million will develop nosocomial infections. Almost 250,000 of these will be bloodstream infections, which contribute significantly to excess length and cost of hospital stays 45 and patient mortality. The attributable mortality from bloodstream infections averages 26 % but varies according to the specific organism involved. [0009] Of all the pathogens isolated, Candida had the highest attributable mortality rate (40 %) [Edmond et al. Clin. Infect. Dis. 29, 239-244, 1999]. Data collected by the NNIS (National Nosocomial Infections Surveillance System) showed that between 1980 and 1989 the incidence of nosocomial candidemia increased by almost 500 % in large teaching 50 hospitals and by 219 % and 370 % in small teaching hospitals and large non-teaching hospitals, respectively [Banerjee et al. Am. J. Med. 91 (suppl. 3B), 86S-89S, 1991]. For an overview of invasiveCandidiasis see, for example, ht- tp://www.doctorfungus.org/mycoses/human/candida/InvasiveOverview.htm. [0010] Invasive fungal infections therefore pose a major challenge for the management of immuno-compromised and other patients. Currently, mortality rates are high and effective treatment is hampered by the lack of reliable early 55 diagnosis. Since the clinical symptoms of IFI are non-specific, with fever often being the only symptom at the outset, there
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