# US 7,740,859 B2 · Polypeptides for the diagnosis and therapy of Leishamaniasis > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,740,859 B2 - Document ID: US7740859B2 - Status: Granted - Kind: B2 - Application number: 10/584,451 - Filed: 2003-12-26 - Published: 2008-02-07 - Granted: 2010-06-22 - Earliest claimed: 2003-12-26 - Assignees: All India Institute of Medical Sciences, Division of Clinicial Microbiology; Department of Biotechnology, Department of Govt of India - Inventors: Sarman Singh; Ramu Sivakumar - Web page: https://patentgrep.com/patent/US7740859B2 ## CPC - C07K 14/44: from protozoa - A61P 33/02: Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis - G01N 33/56905: {Protozoa} - G01N 2469/20: Detection of antibodies in sample from host which are directed against antigens from microorganisms - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract The present invention provides compounds and methods for the detection of anti-leishmanial antibodies in individuals suspected of infection with the protozoan parasite of the genus Leishmania, where the infectious agent is an Indian strain and similar or closely related to Indian Leishmania strains. The compounds provided include polypeptides as shown in SEQ ID NO: 5 or SEQ ID NO: 6 which are useful for the detection of anti-leishmanial antibodies in individuals where the immune responses are elicited against Leishmania species of Indian strains and similar or closely related to Indian Leishmania strains, the compounds are also useful as a vaccine and therapeutic agent to prevent and treat leishmaniasis. The present invention further provides a diagnostic kit consisting of antibody raised against polypeptides as shown in SEQ ID NO: 5 or SEQ ID NO: 6 for detecting leishmanial antigens. ## Claims ### Claim 1 An isolated polypeptide, comprising the amino acid sequence as recited in SEQ ID NO: 5 or SEQ ID NO: 6, wherein said polypeptide has an immunogenic region of a Leishmania antigen. ### Claim 2 (depends on 1) The polypeptide as claimed in claim 1, isolated from a strain of Leishmania donovani. ### Claim 3 A method of detecting anti-leishmanial antibodies in a sample, said method comprising: binding (a) binding anti-leishmanial antibodies from said sample to a polypeptide comprising the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 6 that is bound to a solid support or a carrier; (b) contacting bound antibodies from step (a) with a secondary antibody or a protein, conjugated to an enzyme or a label and that specifically binds to the bound antibodies of step (a); and (c) detecting the anti-leishmanial antibodies in the said sample by detecting the secondary antibody or protein specifically bound to said anti-leishmanial antibodies. ### Claim 4 (depends on 3) The method as claimed in claim 3, wherein the sample is selected from a group consisting of whole blood, serum, plasma and other body fluid. ### Claim 5 (depends on 4) The method as claimed in claim 4, wherein the sample is selected from an animal or a mammal, including human beings. ### Claim 6 (depends on 3) The method as claimed in claim 3, wherein said solid support is selected from a group consisting of nitrocellulose, nylon, latex particles, polypropylene and polystyrene material. ### Claim 7 (depends on 3) The method as claimed in claim 3, wherein said carrier is a gold particle. ### Claim 8 (depends on 3) The method as claimed in claim 3, wherein the anti-human secondary antibody is selected from an antibody classes of IgG, IgM, IgA, IgE and their subclasses. ### Claim 9 (depends on 3) The method as claimed in claim 3, wherein the protein is selected from a group consisting of Protein A and Protein G. ### Claim 10 (depends on 3) The method as claimed in claim 3, wherein the enzyme is selected from the group consisting of Alkaline Phosphatase, Horseradish Peroxidase, β-galactosidase, Urease, Xanthine Oxidase, Glucose Oxidase and penicillinase. ### Claim 11 (depends on 3) The method as claimed in claim 3, wherein said label is selected from a group consisting of enzymes, radioisotopes, biotin, chromophores, fluorophores and chemiluminescent moieties. ### Claim 12 (depends on 3) The method as claimed in claim 3, wherein said detecting step of anti-leishmanial antibodies is selected from the group consisting of detecting fluorescence, detecting chemiluminescence, detecting light absorbance or detecting radio isotopes. ### Claim 13 (depends on 1) A diagnostic kit for detecting anti-leishmanial antibodies comprising a polypeptide as claimed in claim 1, and a secondary antibody or a protein, wherein said secondary antibody or the protein is conjugated to an enzyme or a label, and a reagent for detecting said enzyme or label. ### Claim 14 (depends on 13) The kit as claimed in claim 13, wherein the said polypeptide is bound to a solid support or carrier. ### Claim 15 (depends on 13) The kit as claimed in claim 13, wherein the solid support is selected from the group consisting of nitrocellulose, nylon, latex particles, polypropylene and polystyrene material. ### Claim 16 (depends on 13) The kit as claimed in claim 13, wherein the carrier is a gold particle. ### Claim 17 (depends on 13) The kit as claimed in claim 13, wherein the secondary antibody is selected from an antibody classes of IgG, IgM, IgA, IgE and their subclasses. ### Claim 18 (depends on 13) The kit as claimed in claim 13, wherein the protein is selected from a group consisting of Protein A and Protein G. ### Claim 19 (depends on 13) The kit as claimed in claim 13, wherein said label is selected from a group consisting of an enzyme, a radioisotope, biotin, a chromophore, a fluorophore and a chemiluminescent moiety. ### Claim 20 (depends on 13) The kit as claimed in claim 13, wherein the enzyme is selected from the group consisting of Alkaline Phosphatase, Horseradish Peroxidase, β-galactosidase, Urease, Xanthine Oxidase, Glucose Oxidase and penicillinase. ### Claim 21 (depends on 1) A method of obtaining antibodies to the polypeptide of claim 1, said method comprising injecting said polypeptide into an animal, harvesting the antibodies produced against the polypeptide and purifying said antibodies. ### Claim 22 (depends on 21) The method as claimed in claim 21, wherein the animal is selected from a group consisting of a mouse, a rabbit, a horse, a goat, a sheep, a guinea pig, a pig, a bovine, a rat, a chicken and a hamster. ### Claim 23 (depends on 1) The isolated polypeptide of claim 1 consisting of SEQ ID NO: 5 or SEQ ID NO: 6.