Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis
Prophylaxis with macrocyclic lactone (ML) endectocides is the primary strategy for heartworm control. Recent evidence has confirmed that ML-resistant Dirofilaria immitis isolates have evolved. Comparison of genomes of ML-resistant isolates show they are genetically distinct from wild-type population...
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doaj-28998fc64dd640ada85848cf42f56d212020-11-25T01:53:37ZengElsevierInternational Journal for Parasitology: Drugs and Drug Resistance2211-32072018-12-0183596606Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitisCristina Ballesteros0Cassan N. Pulaski1Catherine Bourguinat2Kathy Keller3Roger K. Prichard4Timothy G. Geary5Institute of Parasitology, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, H9X 3V9, QC, Canada; Corresponding author.School of Veterinary Medicine, Louisiana State University, Skip Bertman Drive, Baton Rouge, LA, 70803, USAInstitute of Parasitology, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, H9X 3V9, QC, CanadaInstitute of Parasitology, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, H9X 3V9, QC, CanadaInstitute of Parasitology, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, H9X 3V9, QC, Canada; Corresponding author.Institute of Parasitology, McGill University, 21111 Lakeshore Road, Sainte Anne de Bellevue, H9X 3V9, QC, Canada; Corresponding author.Prophylaxis with macrocyclic lactone (ML) endectocides is the primary strategy for heartworm control. Recent evidence has confirmed that ML-resistant Dirofilaria immitis isolates have evolved. Comparison of genomes of ML-resistant isolates show they are genetically distinct from wild-type populations. Previously, we identified single nucleotide polymorphisms (SNPs) that are correlated with phenotypic ML resistance. Since reliable in vitro assays are not available to detect ML resistance in L3 or microfilarial stages, the failure to reduce microfilaraemia in infected dogs treated with an ML has been proposed as a surrogate clinical assay for this purpose. The goal of our study was to validate the genotype-phenotype correlation between SNPs associated with ML resistance and failure to reduce microfilaraemia following ML treatment and to identify a minimal number of SNPs that could be used to confirm ML resistance. In this study, 29 participating veterinary clinics received a total of 148 kits containing supplies for blood collection, dosing and prepaid shipping. Patients recruited after a diagnosis of heartworm infection were treated with a single standard dose of Advantage Multi® and a blood sample taken pre- and approximately 2–4 weeks post-treatment. Each sample was processed by performing a modified Knott's Test followed by isolation of microfilariae, genomic DNA extraction and MiSeq sequencing of regions encompassing 10 SNP sites highly correlated with ML resistance. We observed significant correlation of SNP loci frequencies with the ML microfilaricidal response phenotype. Although all predictive SNP combination models performed well, a 2-SNP model was superior to other models tested. The predictive ability of these markers for ML-resistant heartworms should be further evaluated in clinical and epidemiological contexts.http://www.sciencedirect.com/science/article/pii/S2211320718300666 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Cristina Ballesteros Cassan N. Pulaski Catherine Bourguinat Kathy Keller Roger K. Prichard Timothy G. Geary |
spellingShingle |
Cristina Ballesteros Cassan N. Pulaski Catherine Bourguinat Kathy Keller Roger K. Prichard Timothy G. Geary Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis International Journal for Parasitology: Drugs and Drug Resistance |
author_facet |
Cristina Ballesteros Cassan N. Pulaski Catherine Bourguinat Kathy Keller Roger K. Prichard Timothy G. Geary |
author_sort |
Cristina Ballesteros |
title |
Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis |
title_short |
Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis |
title_full |
Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis |
title_fullStr |
Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis |
title_full_unstemmed |
Clinical validation of molecular markers of macrocyclic lactone resistance in Dirofilaria immitis |
title_sort |
clinical validation of molecular markers of macrocyclic lactone resistance in dirofilaria immitis |
publisher |
Elsevier |
series |
International Journal for Parasitology: Drugs and Drug Resistance |
issn |
2211-3207 |
publishDate |
2018-12-01 |
description |
Prophylaxis with macrocyclic lactone (ML) endectocides is the primary strategy for heartworm control. Recent evidence has confirmed that ML-resistant Dirofilaria immitis isolates have evolved. Comparison of genomes of ML-resistant isolates show they are genetically distinct from wild-type populations. Previously, we identified single nucleotide polymorphisms (SNPs) that are correlated with phenotypic ML resistance. Since reliable in vitro assays are not available to detect ML resistance in L3 or microfilarial stages, the failure to reduce microfilaraemia in infected dogs treated with an ML has been proposed as a surrogate clinical assay for this purpose. The goal of our study was to validate the genotype-phenotype correlation between SNPs associated with ML resistance and failure to reduce microfilaraemia following ML treatment and to identify a minimal number of SNPs that could be used to confirm ML resistance. In this study, 29 participating veterinary clinics received a total of 148 kits containing supplies for blood collection, dosing and prepaid shipping. Patients recruited after a diagnosis of heartworm infection were treated with a single standard dose of Advantage Multi® and a blood sample taken pre- and approximately 2–4 weeks post-treatment. Each sample was processed by performing a modified Knott's Test followed by isolation of microfilariae, genomic DNA extraction and MiSeq sequencing of regions encompassing 10 SNP sites highly correlated with ML resistance. We observed significant correlation of SNP loci frequencies with the ML microfilaricidal response phenotype. Although all predictive SNP combination models performed well, a 2-SNP model was superior to other models tested. The predictive ability of these markers for ML-resistant heartworms should be further evaluated in clinical and epidemiological contexts. |
url |
http://www.sciencedirect.com/science/article/pii/S2211320718300666 |
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