Assays to Detect β-Tubulin Codon 200 Polymorphism in Trichuris trichiura and Ascaris lumbricoides
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BACKGROUND: The soil-transmitted helminths (STH) Ascaris lumbricoides and Trichuris trichiura are gastrointestinal parasites causing many disabilities to humans, particularly children. The benzimidazole (BZ) drugs, albendazole (ALB) and mebendazole (MBZ), are commonly used for mass treatment for STH. Unfortunately, there is concern that increased use of anthelmintics could select for resistant populations of these human parasites. In veterinary parasites, and lately in filarial nematodes, a single amino acid substitution from phenylalanine to tyrosine, known to be associated with benzimidazole resistance, has been found in parasite beta-tubulin at position 200. We have developed pyrosequencer assays for codon 200 (TTC or TAC) in A. lumbricoides and T. trichiura to screen for this single nucleotide polymorphism (SNP). METHOD AND FINDINGS: Pyrosequencing assays were developed and evaluated for detecting the TTC or TAC SNP at codon 200 in beta-tubulin in A. lumbricoides and T. trichiura. Genomic DNA from individual worms, eggs isolated from individual adult worms or from fecal samples with known treatment history and origin, were sequenced at beta-tubulin by pyrosequencing, and genotypes were confirmed by conventional sequencing. The assays were applied to adult worms from a benzimidazole-naïve population in Kenya. Following this, these assays were applied to individual worms and pooled eggs from people in East Africa (Uganda and Zanzibar) and Central America (Panama) where mass anthelmintic drug programs had been implemented. All A. lumbricoides samples were TTC. However, we found 0.4% homozygous TAC/TAC in T. trichiura worms from non-treated people in Kenya, and 63% of T. trichiura egg pools from treated people in Panama contained only TAC. CONCLUSION: Although the codon 200 TAC SNP was not found in any of the A. lumbricoides samples analyzed, a rapid genotyping assay has been developed that can be used to examine larger populations of this parasite and to monitor for possible benzimidazole resistance development. The TAC SNP at codon 200, associated with benzimidazole resistance in other nematodes, does occur in T. trichiura, and a rapid assay has been developed to allow populations of this parasite to be monitored for the frequency of this SNP. Sample sizes were small, anthelmintic efficacy was not assessed, and treated and non-treated samples were from different locations, so these frequencies cannot be extrapolated to other populations of T. trichiura or to a conclusion about resistance to treatment. The occurrence of the TAC SNP at codon 200 of beta-tubulin in T. trichiura may explain why benzimidazole anthelmintics are not always highly effective against this species of STH. These assays will be useful in assessing appropriate treatment in areas of high T. trichiura prevalence and in monitoring for possible resistance development in these STH.
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It was suggested that the gene mutation at codon 200 might be related with the drug resistance in T. trichiura.
“It was suggested that the gene mutation at codon 200 might be related with the drug resistance [340,341].”
From Albendazole and Mebendazole as Anti-Parasitic and Anti-Cancer Agents: an Update (2021) · cites this paper as [340]
Diawara et al. confirmed different alleles at codon 200 in the β-tubulin gene from T. trichiura worms (Kenya) and eggs (Panama) obtained from children.
“Diawara et al. [340] confirmed different alleles at codon 200 in the β-tubulin gene (only 1 isotype has been found at this gene in T. trichiura) from T. trichiura worms (Kenya) and eggs (Panama) obtained from children.”
From Albendazole and Mebendazole as Anti-Parasitic and Anti-Cancer Agents: an Update (2021) · cites this paper as [340]
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References 36 · Cited by 135
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- [2]Bundy DAP Shaeffer S Jukes M Beegle K Gillespie A 2006 School-based health and nutrition programs. Jamison DT Breman JG Measham AR Alleyne G Claeson M Disease Control Priorities in Developing Countries. 2nd edition. New York The World Bank and Oxford University Press 1091 1108
- [3]Lacey 1990 Mode of action of benzimidazoles. Parasitol Today 6 112 124 15463312
- [7]Mottier ML Prichard RK 2008 Genetic analysis of a relationship between macrocyclic lactone and benzimidazole anthelmintic selection on Haemonchus contortus. Pharmacogenet Genomics 18 128 140 18192899
- [9]Stothard JR Imison E French MD Sousa-Figueiredo JC Mgeni AF 2008 Soil-transmitted helminthiasis among mothers and their preschool children on Unguja Island, Zanzibar with emphasis upon ascariasis. Parasitology 8 1 9 18775092
- [10]Dryden MW Payne PA Ridley R Smith V 2005 Comparison of common fecal flotation techniques for the recovery of parasites eggs and oocysts. Vet Ther 6 15 28 15906267
- [11]Bennett AB Barker GC Bundy DAP 1999 A beta-tubulin gene from Trichuris trichiura. Mol Biochem Parasitol 103 111 116 10514087
- [12]Nilsen TW Shambaugh J Denker J Chubb G Faser C 1989 Characterization and expression of a spliced leader RNA in the parasitic nematode Ascaris lumbricoides var. suum. Mol Cell Biol 9 3543 3547 2796996 PMC362403
- [13]Kwa MSG Veenstra JG Roos MH 1993 Molecular characterization of β-tubulin genes present in benzimidazole-resistant populations of Haemonchus contortus. Mol Biochem Parasitol 60 133 144 8366887
- [15]Prichard RK 2001 Genetic variability following selection of Haemonchus contortus with anthelmintics. Trends Parasitol 17 445 453 11530357
- [16]Prichard RK 2007 Markers for benzimidazole resistance in human parasitic nematodes? Parasitology 134 1087 1092 17608968
- [17]Prichard RK von Samson-Himmelstjerna G Blackhall WJ Geary T 2007 Foreword: towards markers for anthelmintic resistance in helminths of importance in animal and human health. Parasitology 134 1073 1076 17608966
- [18]Bennett AB Anderson TJC Barker GC Michael E Bundy DAP 2002 Sequence variation in the Trichuris trichiura β-tubulin locus: implications for the development of benzimidazole resistance. Int J Parasitol 32 1519 1528 12392917
- [19]Schwab AE Boakye DA Kyelem D Prichard RK 2005 Detection of benzimidazole resistance-associated mutations in the filarial nematode Wuchereria bancrofti and evidence for selection by albendazole and ivermectin combination treatment. Am J Trop Med Hyg 73 234 238 16103581
- [20]Schwenkenbecher JM Albonico M Bickle Q Kaplan RM 2007 Characterization of beta-tubulin genes in hookworms and investigation of resistance-associated mutations using real-time PCR. Mol Biochem Parasitol 156 167 174 17850900
- [23]Martin PJ Anderson N Jarret RG 1998 Detecting benzimidazole resistance with faecal egg count reduction tests and in vitro assays. Aust Vet J 66 236 240 2590136
- [24]Coles GC Bauer C Borgsteede M Geerts S Klei TR 1992 World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Vet Parasitol 44 35 44 1441190
- [25]Beech RN Prichard RK Scott ME 1994 Genetic variability of the β-tubulin genes in benzimidazole-susceptible and -resistant strains of Haemonchus contortus. Genetics 38 103 110 PMC1206121 8001777
- [26]Elard L Humbert JF 1999 Importance of the mutation of amino acid 200 of the isotype 1 β-tubulin gene in the benzimidazole resistance of a small-ruminant parasite Teladorsagia circumcincta. Parasitol Res 85 452 456 10344538
- [27]Roos MH Boersema JH Borgsteede FH Cornelissen J Taylor M 1990 Molecular analysis of selection for benzimidazole resistance in the sheep parasite Haemonchus contortus. Mol Biochem Parasitol 43 77 88 1981249
- [28]Stephenson LS Latham MC Adams EJ Kioti SN Perter A 1993 Weight gain of Kenyan school children infected with hookworm, Trichuris trichiura and Ascaris lumbricoides is improved following once- or twice-yearly treatment with albendazole. J Nutr 123 656 665 8463866
- [29]Magnussen P Muchiri E Mungai P Ndzovu M Ouma J 1997 A school-based approach to control the urinary schistosomiasis and intestinal helminths infections in children in Matuga, Kenya: impact of a two-year chemotherapy programme on prevalence and intensity of infections. Trop Med Int Health 2 825 831 9315040
- [30]Muchiri EM Thiong'o FW Magnussen P Ouma JH 2001 A comparative study of different albendazole and mebendazole regimens for the treatment of intestinal infections in school children of Usigu Division, western Kenya. J Parasitol 87 413 418 11318574
- [31]Prichard RK 1990 Anthelmintic resistance in nematodes: extent, recent understanding and future directions for control and research. Int J Parasitol 20 515 523 2210945
- [33]Albonico M Bickle Q Ramsan M Montresor A Savioli L 2003 Efficacy of mebendazole and levamisole alone or in combination against intestinal nematode infections after repeated targeted mebendazole treatment in Zanzibar. Bull World Health Organ 81 343 352 12856052 PMC2572452
- [34]Wolstenholme AJ Fairweather I Prichard RK Samson-Himmelstjerna G Sangster NC 2004 Drug resistance in veterinary helminths. Trends Parasitol 20 469 476 15363440
- [35]Adams VJ Lombard CJ Dhansay MA Markus MB Finchman JE 2004 Efficacy of albendazole against whipworm Trichuris trichiura: a randomized, controlled trial. S Afr Med J 94 972 976 15662995
- [36]Pape M Posedi J Failing K Schnieder T von Samson-Himmelstjerna G 2003 Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole-susceptible and -resistant cyathostome population. Parasitology 127 53 59 12885188
- [37]Ghisi M Kaminsky R Maser P 2007 Phenotyping and genotyping of Haemonchus contortus isolates reveals a new putative candidate mutation for benzimidazole resistance in nematodes. Vet Parasitol 144 313 320 17101226
- Authors
- Diawara, A., Drake, L. J., Suswillo, R. R., Kihara, J., Bundy, D. A. P., Scott, M. E., Halpenny, C., Stothard, J. R., & Prichard, R. K.
- Affiliations
- Imperial College London (GB)
- Kenya Medical Research Institute (KE)
- World Bank (US)
- McGill University (CA)
- Natural History Museum (GB)
- Corresponding author
- Prichard, Roger K.
- Venue
- PLoS Neglected Tropical Diseases
- Publisher
- Public Library of Science (PLoS)
- Published
- 2009-03-24
- Volume
- 3
- Issue
- 3
- Pages
- e397
- ISSN
- 1935-2727 (ISSN-L)
- 1935-2735
- PMC
- PMC2654341
- OpenAlex
- W2091868738
- Cited by
- 135
- References
- 36
- Citations by year
- 2026: 3 · 2025: 7 · 2024: 5 · 2023: 4 · 2022: 11 · 2021: 10 · 2020: 10 · 2019: 6 · 2018: 8 · 2017: 6 · 2016: 6 · 2015: 6 · 2014: 15 · 2013: 15 · 2012: 9
- FWCI
- 5.54
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS neglected tropical diseases (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- LSHTM Research Online (London School of Hygiene and Tropical Medicine) (acceptedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Funders
- World Bank Group
- Topics
- Life Sciences › Immunology and Microbiology › Parasitology › Parasites and Host Interactions
- Health Sciences › Veterinary › Small Animals › Helminth infection and control
- Health Sciences › Medicine › Infectious Diseases › Parasitic Diseases Research and Treatment
- Keywords
- Trichuris trichiura, Ascaris lumbricoides, benzimidazole resistance, anthelmintic resistance, soil-transmitted helminths, albendazole, mebendazole, mass drug administration, resistance monitoring, single nucleotide polymorphisms, East Africa
- MeSH terms
- Amino Acid Sequence, Animals, Anthelmintics, Ascariasis, Benzimidazoles, Child, Codon, Drug Resistance, Humans, Kenya, Molecular Sequence Data, Panama, Trichuriasis, Trichuris, Tubulin, Helminth Proteins, Sequence Alignment, Ascaris lumbricoides, Sequence Analysis, DNA, Polymorphism, Single Nucleotide
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