Methods, Devices, Kits and Compositions for Detecting Roundworm, Whipworm, and Hookworm

• US 20140205994A1
• Filed: 03/07/2013
• Published: 07/24/2014
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Monsanto Company (Bayer AG), IDEXX Laboratories Incorporated
• Original Assignee: Divergence, Inc. (Bayer AG), IDEXX Laboratories Incorporated
• Inventors: Elsemore, David Allen, Geng, Jinming, Flynn, Laurie A., Crawford, Michael
• Granted Patent: US 8,895,294 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: G01N 2333/43526 from worms
• CPC Class Codes: G01N 2333/4353 from nematodes
• CPC Class Codes: G01N 2469/10 Detection of antigens from microorganism in sample from host
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Methods, devices, kits and compositions for detecting the presence or absence of one or more helminthic coproantigens in a sample are disclosed herein. The methods, devices, kits and compositions of the present invention may be used to confirm the presence or absence of roundworm, whipworm and/or hookworm in a fecal sample from a mammal and may also be able to distinguish between one or more helminth infections. Confirmation of the presence or absence of roundworm, whipworm and/or 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

1-61. -61. (canceled)

62. A method of detecting the presence or absence of one or more helminthic antigens in a sample, the method comprising:

(a) contacting a sample from a mammal with two antibodies selected from the group consisting of;

(i) a first antibody capable of specifically binding a roundworm coproantigen, but not a whipworm coproantigen and a second antibody capable of specifically binding a whipworm coproantigen, but not a roundworm coproantigen;

(ii) a first antibody capable of specifically binding a roundworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a roundworm coproantigen; and

(iii) a first antibody capable of specifically binding a whipworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a whipworm coproantigen;

(b) forming antibody-coproantigen complexes in the presence of the coproantigens, if any, in the sample; and

(c) detecting the presence or absence of the antibody-coproantigen complexes, if any.

64. The method according to claim 62 or 63, 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-worm parasites, heartworm, one or more viruses, one or more fungi, and one or more bacteria.

65. The method according to claim 62 or 63, wherein the reagents for the detection of any one or all of the 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-worm parasites, one or more viruses, one or more fungi or one or more bacteria.

66. The method according to claim 62 or 63, wherein the antibody capable of specifically binding a roundworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

38, or Copro6728;

or the first antibody was raised against an extract of whole roundworms, or extract of roundworm reproductive organs, or extract of roundworm intestines.

67. The method according to claim 62 or 63, wherein the antibody capable of specifically binding a whipworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

9.

68. The method according to claim 62 or 63, wherein the antibody capable of specifically binding a hookworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

33 or SEQ ID NO;

34, or CoproASP5.

69. The method according to claim 62 or 63, wherein the sample is obtained from a mammal that is a canine or a feline.

70. The method according to claim 62 or 63, wherein the roundworm is Toxocara canis or Toxocara cati, Toxocara vitulorum, Toxascaris leonina, Baylisascaris procyonis, Ascaridia galli, Parascaris equorum, Ascaris suum, Ascaris lumbricoides, Anisakis simplex, or Pseudoterranova decipiens.

71. The method according to claim 62 or 63, wherein the whipworm is Trichuris vulpis, Trichuris campanula, Trichuris serrata, Trichuris suis, Trichuris trichiura, Trichuris discolor and Trichocephalus trichiuris.

72. The method according to claim 62 or 63, wherein the hookworm is Ancylostoma caninum, Ancylostoma braziliense, Ancylostoma duodenal, Ancylostoma ceylanicum, Ancylostoma tubaeforme and Ancylostoma pluridentatum, Necator americanus, and Uncinaria stenocephala.

73. The method according to claim 62 or 63, wherein the first and second antibodies do not specifically bind any coproantigen derived from heartworm.

74. The method according to claim 62 or 63, wherein the step of detecting the presence or absence of the complexes further includes the step of providing at least one secondary antibody that binds to at least one of the complexes.

75. The method according to claim 74, wherein the at least one secondary antibody is labeled.

76. The method according to claim 62 or 63, wherein one or more of the first and second antibodies are labeled.

77. The method according to claim 62 or 63, wherein the first and second antibodies are immobilized on a solid support.

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

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

80. The method according to claim 62 or 63, wherein the roundworm coproantigen, hookworm coproantigen, and/or whipworm coproantigen is from a fecal sample.

63. A method of diagnosing whether a mammal is infected with one or more parasitic worms, the method comprising the steps of:

(a) contacting a sample from a mammal with two antibodies selected from the group consisting of;

(i) a first antibody capable of specifically binding a roundworm coproantigen, but not a whipworm coproantigen and a second antibody capable of specifically binding a whipworm coproantigen, but not a roundworm coproantigen;

(ii) a first antibody capable of specifically binding a roundworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a roundworm coproantigen; and

(iii) a first antibody capable of specifically binding a whipworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a whipworm coproantigen;

(b) forming antibody-coproantigen complexes in the presence of the coproantigens, if any, in the sample;

(c) detecting the presence or absence of the antibody-coproantigen complexes, if any; and

(d) diagnosing the mammal as having;

(i) a roundworm infection if a roundworm antibody-coproantigen complex is present;

(ii) a whipworm infection if a whipworm antibody-coproantigen complex is present; and

(iii) a hookworm infection if a hookworm antibody-coproantigen complex is present.

81. A device for detecting the presence or absence of one or more helminthic antigens from a sample, the device comprising a solid support, wherein the solid support has immobilized thereon two antibodies selected from the group consisting of:

(i) a first antibody capable of specifically binding a roundworm coproantigen, but not a whipworm coproantigen and a second antibody capable of specifically binding a whipworm coproantigen, but not a roundworm coproantigen;

(ii) a first antibody capable of specifically binding a roundworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a roundworm coproantigen; and

(iii) a first antibody capable of specifically binding a whipworm coproantigen, but not a hookworm coproantigen and a second antibody capable of specifically binding a hookworm coproantigen, but not a whipworm coproantigen.

82. The device according to claim 81, further comprising one or more reagents to detect one or more of the group consisting of:

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

83. The device according to claim 81, wherein said one or more reagents are one or more antibodies or one or more antigens recognized by antibodies specific for one or more non-worm parasites, one or more viruses, one or more fungi or one or more bacteria.

84. The device according to claim 81, wherein the antibody capable of specifically binding a roundworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

26, SEQ ID NO;

27, SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, SEQ ID NO;

38, or Copro6728;

or the first antibody was raised against an extract of whole roundworms, or extract of roundworm reproductive organs, or extract of roundworm intestines.

85. The device according to claim 81, wherein the antibody capable of specifically binding a whipworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, or SEQ ID NO;

9.

86. The device according to claim 81, wherein the antibody capable of specifically binding a hookworm coproantigen is capable of specifically binding a polypeptide consisting of an amino acid sequence of SEQ ID NO:

33 or SEQ ID NO;

34, or CoproASP5.

87. The device according to claim 81, wherein the sample is obtained from a mammal that is a canine or a feline.

88. The device according to claim 81, wherein the roundworm is Toxocara canis or Toxocara cati, Toxocara vitulorum, Toxascaris leonina, Baylisascaris procyonis, Ascaridia galli, Parascaris equorum, Ascaris suum, Ascaris lumbricoides, Anisakis simplex, or Pseudoterranova decipiens.

89. The device according to claim 81, wherein the whipworm is Trichuris vulpis, Trichuris campanula, Trichuris serrata, Trichuris suis, Trichuris trichiura, Trichuris discolor and Trichocephalus trichiuris.

90. The device according to claim 81, wherein the hookworm is Ancylostoma caninum, Ancylostoma braziliense, Ancylostoma duodenal, Ancylostoma ceylanicum, Ancylostoma tubaeforme and Ancylostoma pluridentatum, Necator americanus, and Uncinaria stenocephala.

91. The device according to claim 81, wherein the first and second antibodies do not specifically bind any coproantigen derived from heartworm.

92. The device according to claim 81, wherein the step of detecting the presence or absence of the complexes further includes the step of providing at least one secondary antibody that binds to at least one of the complexes.

93. The device according to claim 92, wherein the at least one secondary antibody is labeled.

94. The device according to claim 81, wherein one or more of the first and second antibodies are labeled.

95. The device according to claim 81, wherein the solid support forms part of an enzyme-linked immunosorbent assay device.

96. The device according to claim 95, wherein the enzyme-linked immunosorbent assay device is a lateral flow immunoassay device.

97. The device according to claim 81, wherein the roundworm coproantigen, hookworm coproantigen, and/or whipworm coproantigen is from a fecal sample.

98. A kit for detection of one or more roundworm coproantigens, one or more whipworm coproantigens and/or one or more hookworm coproantigens in a mammalian sample, the kit comprising the device of claim 81, and one or more reagents sufficient for the detection of the one or more antigens.

99. The kit according to claim 98, wherein the one or more reagents are selected from the group consisting of one or more indicator reagents, one or more antibody labeling compounds, one or more antibodies, one or more antigen capture reagents, one or more inhibitors, and one or more wash reagents.