Macrocyclic lactone resistance marker for
View Patent ↗The invention relates to a method for determining the responsiveness of a Dirofilaria spp. nematode to a macrocyclic lactone, by determining the genotype of the nematode at a position in a P-glycoprotein gene of the nematode corresponding to position 11, and optionally to position 618, in SEQ ID NO: 1, wherein the genotype GG at a position corresponding to position 11, or at positions corresponding to positions 11 and 618, in SEQ ID NO: 1 indicates that the nematode is likely resistant to the macrocyclic lactone. The invention also relates to methods for selecting a treatment to treat an animal infected with macrocyclic lactone resistant Dirofilaria spp nematode, and treating the animal; or for selecting a prophylactic to prevent an animal from becoming infected with a macrocyclic lactone resistant Dirofilaria spp nematode, and providing the prophylactic to the animal. The invention further relates to an isolated nucleic acid having at least 80% sequence identity to SEQ ID NO: 1.
1. A kit for determining the responsiveness of a Dirofilaria spp. nematode to a macrocyclic lactone, the kit comprising: a probe capable of determining the genotype of the Dirofilaria spp. nematode at a position in a P-glycoprotein gene of the nematode corresponding to position 11 of SEQ ID NO: 1 or a nucleotide sequence at least 80% identical to SEQ ID NO: 1; or a probe capable of determining the genotype of the Dirofilaria spp. nematode at positions in a P-glycoprotein gene of the nematode corresponding to positions 11 and 618 of SEQ ID NO: 1; wherein said probe comprises a label and wherein the genotype GG at said position corresponding to position 11 in SEQ ID NO: 1 or the genotype GG at said positions corresponding to positions 11 and 618 in SEQ ID NO: 1, indicates that the nematode is likely to be resistant to said macrocyclic lactone.
2. The kit according to claim 1 , wherein said Dirofilaria spp. nematode is Dirofilaria immitis.
3. The kit according to claim 1 , wherein the macrocyclic lactone is selected from the group consisting of ivermectin, selamectin, milbemycin oxime and moxidectin.
4. The kit according to claim 1 , wherein the probe is an oligonucleotide, a primer, an aptamer or an antibody.
5. A kit according to claim 1 wherein the label is a radioactive, enzymatic or fluorescent tag.
6. A kit according to claim 1 further comprising one or more reagents, buffers, packaging materials, instructions for using the kit or containers for holding components of the kit.
7. A method for treating an animal infected with a Dirofilaria spp. nematode, the method comprising determining for an individual or population of Dirofilaria spa nematodes in an area, the genotype of said individual or population of nematodes at a position in a P-glycoprotein gene of said individual or population of nematodes corresponding to position 11 in SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1, or corresponding to positions 11 and 618 in SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1, wherein said determining comprises the use of a probe which binds to a nucleotide sequence corresponding to position 11 in SEQ ID NO:1, or a nucleotide sequence at least 80% identical to SEQ ID NO:1 or which binds to a nucleotide sequence corresponding to positions 11 and 618 of SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1; or which binds to a position in P-glycoprotein corresponding to position 11 of SEQ ID NO:1, or a sequence at least 80% identical to SEQ ID NO:1 or a position in P-glycoprotein corresponding to position 11 and 618 of SEQ ID NO:1, or a nucleotide sequence at least 80% identical to SEQ ID NO:1 and administering to said animal an adjusted dosage of a macrocyclic lactone, or administering an arsenic therapy, diethylcarbamazine, antibiotics, or a combination of one or more thereof, if the individual or population of nematodes has the genotype GG at a position in the P-glycoprotein gene corresponding to position 11 in SEQ ID NO: 1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1; or corresponding to positions 11 and 618 in SEQ ID NO:1, or in a nucleotide sequence at least 80% identical to SEQ ID NO:1.
8. The method according to claim 7 , wherein the Dirofilaria spp. nematode is Dirofilaria immitis.
9. The method of claim 7 wherein the arsenic therapy is melarsomine dihydrochloride.
10. The method of claim 7 wherein the macrocyclic lactone is selected from the group consisting of ivermectin, selamectin, milbemycin oxime and moxidectin.
11. The method of claim 7 wherein the antibiotic is tetracycline.
12. The method of claim 11 wherein the tetracycline is doxycycline.
13. A method for treating an animal infected with a Dirofilaria spp. nematode, the method comprising: determining the genotype of said nematode at a position in a P-glycoprotein gene of said nematode corresponding to position 11 in SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1, or corresponding to positions 11 and 618 in SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1, wherein said determining comprises the use of a probe which binds to a nucleotide sequence corresponding to position 11 in SEQ ID NO:1, or a nucleotide sequence at least 80% identical to SEQ ID NO:1 or which binds to a nucleotide sequence corresponding to positions 11 and 618 of SEQ ID NO:1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1; or which binds to a position in P-glycoprotein corresponding to position 11 of SEQ ID NO:1, or a sequence at least 80% identical to SEQ ID NO:1 or a position in P-glycoprotein corresponding to position 11 and 618 of SEQ ID NO:1, or a nucleotide sequence at least 80% identical to SEQ ID NO:1 and administering to said animal an adjusted dosage of a macrocyclic lactone, or administering an arsenic therapy, diethylcarbamazine, antibiotics, or a combination of one or more thereof, if the nematode of said infected animal has the genotype GG at a position in the P-glycoprotein gene corresponding to position 11 in SEQ ID NO: 1 or a nucleotide sequence at least 80% identical to SEQ ID NO:1; or corresponding to positions 11 and 618 in SEQ ID NO:1, or in a nucleotide sequence at least 80% identical to SEQ ID NO:1.
14. The method of claim 13 wherein said Dirofilaria spp. nematode is Dirofilaria immitis.
15. The method of claim 13 wherein the macrocyclic lactone is selected from the group consisting of ivermectin, selamectin, milbemycin oxime and moxidectin.
16. The method of claim 13 wherein the arsenic therapy is melarsomine dihydrochloride.
17. The method of claim 13 wherein the antibiotic is tetracycline.
18. The method of claim 17 wherein the tetracycline is doxycycline.