Compositions, Devices, Kits and Methods for Detecting Hookworm

• US 20100151500A1
• Filed: 05/18/2009
• Published: 06/17/2010
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: IDEXX Laboratories Incorporated
• Original Assignee: IDEXX Laboratories Incorporated
• Inventors: Geng, Jinming, Elsemore, David Allen
• Granted Patent: US 9,239,326 B2
• Time in Patent Office: Days
• US Class Current: 435/7.220
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: G01N 2333/43526 from worms
• CPC Class Codes: G01N 2333/4353 from nematodes
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Abstract

Methods, devices, kits and compositions for detecting the presence or absence of hookworm in a fecal sample are disclosed herein. The methods, devices, kits and compositions of the present invention may be used to confirm the presence or absence of hookworm in a fecal sample from a mammal that may also be infected with one or more of roundworm, whipworm, and heartworm. Confirmation of the presence or absence of hookworm in the mammal may be made, for example, for the purpose of selecting an optimal course of treating the mammal and/or for the purpose of determining whether the mammal has been rid of the infection after treatment has been initiated.

Claims

1. A purified CoproASP5 polypeptide.

52. A method of detecting the presence or absence of one or more hookworm antigens in a sample comprising:

(a) contacting the sample with one or more antibodies that can specifically bind to the polypeptide of claim 1; and

(b) detecting the presence or absence of the one or more hookworm antigens or detecting the presence or absence of one or more complexes that include the one or more hookworm antigens and the one or more antibodies.

53. The method of claim 52 wherein the one or more antibodies are obtained by immunization with a polypeptide that has an amino acid sequence that corresponds to SEQ ID NO:

4, or an antigenic portion thereof,

54. The method of claim 52 wherein the amino acid sequence of the polypeptide has 30 or fewer conserved amino acid substitutions to SEQ ID NO:

4.

55. The method of claim 52 wherein the amino acid sequence of the polypeptide is a conservative variant of SEQ ID NO:

4.

56. The method of claim 52 wherein the sample is obtained from a mammal that is a canine.

57. The method of claim 52 wherein the hookworm is Ancylostoma caninum or Ancylostoma braziliense.

58. The method of claim 52 wherein the sample is a fecal sample.

59. The method of claim 52 wherein the one or more antibodies do not specifically bind any coproantigen derived from the group consisting of roundworm, whipworm, and heartworm.

60. The method of claim 52 wherein the step of detecting the presence or absence of the one or more complexes further includes the step of providing a secondary antibody that binds to the one or more complexes.

61. The method of claim 52 wherein the secondary antibody is labeled.

62. The method of claim 61 wherein one or more of the one or more antibodies are labeled.

63. The method of claim 52 wherein the one or more antibodies are immobilized on a solid support.

64. The method of claim 63 wherein the solid support forms part of an enzyme-linked immunosorbent assay device.

65. The method of claim 64 wherein the enzyme-linked immunosorbent assay device is a lateral flow immunoassay device.

66. The method of claim 52 further comprising the step of contacting the sample with one or more reagents to detect one or more of the group consisting of:

one or more non-hookworm worm parasites, one or more non-worm parasites, one or more viruses, one or more fungi, and one or more bacteria.

67. The method of claim 66 wherein the reagents for the detection of any one or all of the one or more non-hookworm worm parasites, one or more non-worm parasites, one or more viruses, one or more fungi and one or more bacteria are one or more antibodies or one or more antigens recognized by antibodies specific for the one or more non-hookworm worm parasites, one or more non-worm parasites, one or more viruses, one or more fungi or one or more bacteria.

68. A method of diagnosing an intestinal hookworm infection in a mammal, the method comprising the steps of:

(a) obtaining a fecal sample from the mammal

(b) contacting the sample from the mammal with one or more antibodies that can specifically bind to the polypeptide of claim 1;

(c) detecting the presence or absence of the one or more hookworm antigens or detecting the presence or absence of one or more complexes that include the one or more hookworm antigens and the one or more antibodies; and

(d) diagnosing the mammal as having or as not having the hookworm infection based on the detection of the presence or absence of the one or more polypeptides or the one or more complexes.

69. The method of claim 68 wherein the one or more antibodies are obtained by immunization with a polypeptide that has an amino acid sequence that corresponds to SEQ ID NO:

4, or an antigenic portion thereof,

70. The method of claim 69 wherein the amino acid sequence of the polypeptide has 30 or fewer conserved amino acid substitutions to SEQ ID NO:

4.

71. The method of claim 70 wherein the amino acid sequence of the polypeptide is a conservative variant of SEQ ID NO:

4.

72. The method of claim 68 wherein the mammal is a canine or a feline.

73. The method of claim 68 wherein the hookworm is Ancylostoma caninum.

74. The method of claim 68 wherein the step of detecting the presence or absence of the one or more complexes further includes the step of providing a secondary antibody that binds to the one or more complexes.

75. The method of claim 74 wherein the secondary antibody is labeled.

76. The method of claim 68 wherein one or more of the one or more antibodies are labeled.

77. The method of claim 68 wherein the one or more antibodies are immobilized on a solid support.

78. The method of claim 77 wherein the solid support forms part of an enzyme-linked immunosorbent assay device.

79. The method of claim 78 wherein the enzyme-linked immunosorbent assay device is a lateral flow immunoassay device.

80. The method of claim 68 further comprising the step of contacting the sample with one or more reagents to detect one or more of the group consisting of:

one or more non-hookworm worm parasites, one or more-non-worm parasites, one or more viruses, one or more fungi, and one or more bacteria.

81. The method of claim 80 wherein the reagents for the detection of any one or all of the one or more non-hookworm worm parasites, one or more non-worm parasites, one or more viruses, one or more fungi and the one or more bacteria are one or more antibodies or one or more antigens recognized by antibodies specific for one or more non-hookworm worm parasites, one or more non-worm parasites, one or more viruses, one or more fungi or one or more bacteria.

2-51. -51. (canceled)