Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment
Concerns about the emergence of benzimidazole resistance in soil-transmitted helminths (STH) infections, particularly against Trichuris trichiura, have arisen. Previous studies of veterinary nematodes have linked benzimidazole resistance to single-nucleotide polymorphisms (SNPs) at three specific co...
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Elsevier
2024-12-01
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| Series: | International Journal for Parasitology: Drugs and Drug Resistance |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211320724000514 |
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| author | Javier Gandasegui Berta Grau-Pujol Valdemiro Novela Osvaldo Muchisse Maria Cambra-Pellejà Anélsio Cossa José Carlos Jamine Charfudin Sacoor Eric A.T. Brienen Francesc Catala-Moll Lisette van Lieshout María Martínez-Valladares Roger Paredes José Muñoz Stephen R. Doyle |
| author_facet | Javier Gandasegui Berta Grau-Pujol Valdemiro Novela Osvaldo Muchisse Maria Cambra-Pellejà Anélsio Cossa José Carlos Jamine Charfudin Sacoor Eric A.T. Brienen Francesc Catala-Moll Lisette van Lieshout María Martínez-Valladares Roger Paredes José Muñoz Stephen R. Doyle |
| author_sort | Javier Gandasegui |
| collection | DOAJ |
| description | Concerns about the emergence of benzimidazole resistance in soil-transmitted helminths (STH) infections, particularly against Trichuris trichiura, have arisen. Previous studies of veterinary nematodes have linked benzimidazole resistance to single-nucleotide polymorphisms (SNPs) at three specific codons in the beta-tubulin gene, but similar associations in STH have not been consistently observed. In this work, we screened the complete beta-tubulin gene previously linked to benzimidazole resistance in T. trichiura by deep-amplicon sequencing to identify genetic variants and associate levels of diversity with drug response to albendazole. We used 99 DNA samples extracted from T. trichiura pooled eggs, previously semi-purified from human stool samples collected in Manhiça district, Mozambique. We obtained a set of 39 amplicons of the complete gene by subjecting the pooled eggs to long-read PCR and subsequently sequencing them. Of those amplicons, 22 and 17 were obtained from stool samples collected before, and 21 days after albendazole treatment, respectively. We observed genetic variation across the whole gene sequence, in both exons and introns; however, none were associated with the previously proposed resistance-associated SNPs, and none were predicted to significantly affect protein function. No significant differences in genetic diversity were observed between pre- and post-treatment samples. Using publicly available genome-wide data, we also analysed a second beta-tubulin isotype in the T. trichiura genome. We focused on detecting the canonical SNPs and assessing for signatures of genetic selection around this second isotype gene. This analysis did not reveal evidence supporting this second isotype's role in anthelmintic resistance. Despite the limitations of our study, such as a small sample size, particularly paired pre- and post-treatment samples (n = 6), or a restricted geographical area, we found no evidence linking either of the two beta-tubulin genes to benzimidazole resistance in T. trichiura, suggesting that genetic markers of drug resistance likely exist outside the beta-tubulin genes. |
| format | Article |
| id | doaj-art-8dd28f53533c4d828d9a73084451fdce |
| institution | Kabale University |
| issn | 2211-3207 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
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| series | International Journal for Parasitology: Drugs and Drug Resistance |
| spelling | doaj-art-8dd28f53533c4d828d9a73084451fdce2024-12-12T05:21:14ZengElsevierInternational Journal for Parasitology: Drugs and Drug Resistance2211-32072024-12-0126100570Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatmentJavier Gandasegui0Berta Grau-Pujol1Valdemiro Novela2Osvaldo Muchisse3Maria Cambra-Pellejà4Anélsio Cossa5José Carlos Jamine6Charfudin Sacoor7Eric A.T. Brienen8Francesc Catala-Moll9Lisette van Lieshout10María Martínez-Valladares11Roger Paredes12José Muñoz13Stephen R. Doyle14Wellcome Sanger Institute, Cambridgeshire, United Kingdom; Barcelona Institute for Global Health (ISGlobal), Hospital Clínic - University of Barcelona, Barcelona, Spain; Corresponding author. Wellcome Sanger Institute, Cambridgeshire, CB10 1SA, USA.Barcelona Institute for Global Health (ISGlobal), Hospital Clínic - University of Barcelona, Barcelona, Spain; Manhiça Health Research Centre (CISM), Maputo, Mozambique; Mundo Sano Foundation, Buenos Aires, ArgentinaManhiça Health Research Centre (CISM), Maputo, MozambiqueManhiça Health Research Centre (CISM), Maputo, MozambiqueBarcelona Institute for Global Health (ISGlobal), Hospital Clínic - University of Barcelona, Barcelona, Spain; GraphenicaLab S.L., Barcelona, Spain; Facultat de Medicina I Ciències de La Salut, Universitat de Barcelona (UB), Barcelona, SpainManhiça Health Research Centre (CISM), Maputo, MozambiqueManhiça Health Research Centre (CISM), Maputo, MozambiqueManhiça Health Research Centre (CISM), Maputo, MozambiqueLeiden University Center for Infectious Diseases. Parasitology Research Group, Leiden University Medical Center (LUMC), the NetherlandsIrsiCaixa, Badalona, SpainLeiden University Center for Infectious Diseases. Parasitology Research Group, Leiden University Medical Center (LUMC), the NetherlandsInstituto de Ganadería de Montaña (CSIC-Universidad de León), León, SpainIrsiCaixa, Badalona, Spain; Infectious Disease Networking Biomedical Research Center, Centro de Investigación Biomédica en Red de Enfermedades Infecciosas (CIBERINFEC), Carlos III Health Institute, Madrid, Spain; Universitat de Vic-Universitat Central de Catalunya, Vic, Spain; Department of Infectious Diseases, Hospital Universitari Germans Trias I Pujol, Badalona, Spain; Center for Global Health and Diseases, Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH, USABarcelona Institute for Global Health (ISGlobal), Hospital Clínic - University of Barcelona, Barcelona, SpainWellcome Sanger Institute, Cambridgeshire, United KingdomConcerns about the emergence of benzimidazole resistance in soil-transmitted helminths (STH) infections, particularly against Trichuris trichiura, have arisen. Previous studies of veterinary nematodes have linked benzimidazole resistance to single-nucleotide polymorphisms (SNPs) at three specific codons in the beta-tubulin gene, but similar associations in STH have not been consistently observed. In this work, we screened the complete beta-tubulin gene previously linked to benzimidazole resistance in T. trichiura by deep-amplicon sequencing to identify genetic variants and associate levels of diversity with drug response to albendazole. We used 99 DNA samples extracted from T. trichiura pooled eggs, previously semi-purified from human stool samples collected in Manhiça district, Mozambique. We obtained a set of 39 amplicons of the complete gene by subjecting the pooled eggs to long-read PCR and subsequently sequencing them. Of those amplicons, 22 and 17 were obtained from stool samples collected before, and 21 days after albendazole treatment, respectively. We observed genetic variation across the whole gene sequence, in both exons and introns; however, none were associated with the previously proposed resistance-associated SNPs, and none were predicted to significantly affect protein function. No significant differences in genetic diversity were observed between pre- and post-treatment samples. Using publicly available genome-wide data, we also analysed a second beta-tubulin isotype in the T. trichiura genome. We focused on detecting the canonical SNPs and assessing for signatures of genetic selection around this second isotype gene. This analysis did not reveal evidence supporting this second isotype's role in anthelmintic resistance. Despite the limitations of our study, such as a small sample size, particularly paired pre- and post-treatment samples (n = 6), or a restricted geographical area, we found no evidence linking either of the two beta-tubulin genes to benzimidazole resistance in T. trichiura, suggesting that genetic markers of drug resistance likely exist outside the beta-tubulin genes.http://www.sciencedirect.com/science/article/pii/S2211320724000514Trichuris trichiuraDeep-amplicon sequencingBeta-tubulinBenzimidazole resistanceGenomics |
| spellingShingle | Javier Gandasegui Berta Grau-Pujol Valdemiro Novela Osvaldo Muchisse Maria Cambra-Pellejà Anélsio Cossa José Carlos Jamine Charfudin Sacoor Eric A.T. Brienen Francesc Catala-Moll Lisette van Lieshout María Martínez-Valladares Roger Paredes José Muñoz Stephen R. Doyle Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment International Journal for Parasitology: Drugs and Drug Resistance Trichuris trichiura Deep-amplicon sequencing Beta-tubulin Benzimidazole resistance Genomics |
| title | Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment |
| title_full | Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment |
| title_fullStr | Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment |
| title_full_unstemmed | Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment |
| title_short | Deep-amplicon sequencing of the complete beta-tubulin gene in Trichuris trichiura before and after albendazole treatment |
| title_sort | deep amplicon sequencing of the complete beta tubulin gene in trichuris trichiura before and after albendazole treatment |
| topic | Trichuris trichiura Deep-amplicon sequencing Beta-tubulin Benzimidazole resistance Genomics |
| url | http://www.sciencedirect.com/science/article/pii/S2211320724000514 |
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