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Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions

  • Fattaneh Mikaeili , Alexander Mathis , Peter Deplazes , Hossein Mirhendi , Afshin Barazesh , Sepideh Ebrahimi and Eshrat Beigom Kia EMAIL logo
Published/Copyright: July 5, 2017
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Acta Parasitologica
From the journal Acta Parasitologica Volume 62 Issue 3

Abstract

The definitive genetic identification of Toxocara species is currently based on PCR/sequencing. The objectives of the present study were to design and conduct an in silico polymerase chain reaction-restriction fragment length polymorphism method for identification of Toxocara species. In silico analyses using the DNASIS and NEBcutter softwares were performed with rDNA internal transcribed spacers, and mitochondrial cox1 and nad1 sequences obtained in our previous studies along with relevant sequences deposited in GenBank. Consequently, RFLP profiles were designed and all isolates of T. canis and T. cati collected from dogs and cats in different geographical areas of Iran were investigated with the RFLP method using some of the identified suitable enzymes. The findings of in silico analyses predicted that on the cox1 gene only the MboII enzyme is appropriate for PCR-RFLP to reliably distinguish the two species. No suitable enzyme for PCR-RFLP on the nad1 gene was identified that yields the same pattern for all isolates of a species. DNASIS software showed that there are 241 suitable restriction enzymes for the differentiation of T. canis from T. cati based on ITS sequences. RsaI, MvaI and SalI enzymes were selected to evaluate the reliability of the in silico PCR-RFLP. The sizes of restriction fragments obtained by PCR-RFLP of all samples consistently matched the expected RFLP patterns. The ITS sequences are usually conserved and the PCR-RFLP approach targeting the ITS sequence is recommended for the molecular differentiation of Toxocara species and can provide a reliable tool for identification purposes particularly at the larval and egg stages.

Keywords: PCR-RFLP; Toxocara canis; Toxocara cati; internal transcribed spacers (ITS); identification

Acknowledgements

The authors would like to thank Jeannine Hauri, laboratory technician of the vector entomology unit at the institute of parasitology, University of Zurich for kind assistance. This research has been partly supported by the Tehran University of Medical Sciences (Project No. 90-02-27-14163) and Shiraz University of Medical Sciences (Project No. 93-7422). The authors declare that there is no conflict of interests.

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Received: 2016-7-26
Revised: 2017-3-2
Accepted: 2017-4-7
Published Online: 2017-7-5
Published in Print: 2017-9-26

© 2017 W. Stefański Institute of Parasitology, PAS

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  50. Erratum to: 2017, 62(1) should be 2017, 62(2)
https://doi.org/10.1515/ap-2017-0066
Keywords for this article
PCR-RFLP; Toxocara canis; Toxocara cati; internal transcribed spacers (ITS); identification

Articles in the same Issue

  1. Review
  2. Snail species diversity impacts the infection patterns of Echinostoma spp.: Examples from field collected data
  4. Molecular phylogeny of the Haplosplanchnata Olson, Cribb, Tkach, Bray and Littlewood, 2003, with a description of Schikhobalotrema huffmani n. sp.
  6. Structuraland antigenic analysis of a new Rhoptry Pseudokinase Gene (ROP54) in Toxoplasma gondii
  8. Ultrastructure of the spermatozoon of Macvicaria obovata (Digenea: Opecoelidae), a parasite of Sparus aurata(Pisces: Teleostei) from the Gulf of Gabès, Mediterranean Sea
  10. Epidemiology of Toxoplasma gondii infection in native Tibetans in Tibet, China
  12. A PCR survey of vector-borne pathogens in different dog populations from Turkey
  14. Diplectanids (Monogenea) parasitic on sciaenid fish from Peru with the proposal of Pseudorhamnocercoides n. gen., the description of Rhamnocercus dominguesi n. sp. and the redescription of Rhamnocercoides menticirrhi Luque and Iannacone, 1991
  16. Differentiation of Toxocara canis and Toxocara cati based on PCR-RFLP analyses of rDNA-ITS and mitochondrial cox1 and nad1 regions
  18. The Prevalence of Cyclospora cayetanensis and Cryptosporidium spp. in Turkish patients infected with HIV-1
  20. Molecular characterization and functional analysis of a glutathione peroxidase gene from Aphelenchoides besseyi (Nematoda: Aphelenchoididae)
  22. Durettenema guangdongense gen. et sp. nov. (Nematoda: Molineoidea) from Hipposideros larvatus (Horsfield) (Chiroptera: Rhinolophidae) with discussion of the taxonomic status of Macielia rhinolophi Yin, 1980
  24. In vitro effects of purine and pyrimidine analogues on Leishmania donovani and Leishmania infantum promastigotes and intracellular amastigotes
  26. Population structure of the monogenean, Paramazocraes thrissocles (Mazocraeidae) on the gills of clupeoid fish, Thryssa mystax of Visakhapatnam coast, Bay of Bengal
  28. Redescription of Eutarsopolipus elongatus Regenfuss, 1968 (Acari: Podapolipidae) parasitising carabid beetles, with first description of the male
  30. Seroprevalence and risk factors of Neospora spp. and Toxoplasma gondii infections among horses and donkeys in Nigeria, West Africa
  32. Spinicauda sumatrana sp. nov. (Nematoda: Heterakidae) from Ludeking’s Crested Dragon, Lophocalotes ludekingi (Agamidae), from the Bukit Barisan Range of Sumatra
  34. The systematic position and structure of the genus Leyogonimus Ginetsinskaya, 1948 (Platyhelminthes: Digenea) with comments on the taxonomy of the superfamily Microphalloidea Ward, 1901
  36. Leishmania infantum is present in vaginal secretions of naturally infected bitches at lower levels in oestrogenized bitches than in non-oestrogenized bitches
  38. Intestinal coccidian parasites as an underestimated cause of travellers’ diarrhoea in Polish immunocompetent patients
  40. Hymenolepididae (Cestoda, Cyclophyllidea) of ciconiiform and pelecaniform birds: the genera Oschmarinolepis Spassky and Spasskaja, 1954 and Paraoschmarinolepis gen. n.
  42. Redescription of two species of Thaparocleidus (Monogenea: Dactylogyridae), with the description of T. armillatus sp. n. from Wallago attu and a phylogenetic analysis based on 18S rDNA sequences
  44. In vitro and in vivo studies of nematophagous fungi Arthrobotrys musiformis and Arthrobotrys robusta against the larvae of the trichostrongylides
  46. Three monogeneans parasitic on marine sciaenid fish from Peru including description of Cynoscionicola veranoi n. sp. (Microcotylidae), and redescription of C. americanus Tantaleán, Martínez and Escalante, 1987 and Hargicotyle sciaenae Oliva and Luque, 1989 (Diclidophoridae)
  48. New and previously described dactylogyrid species (Monogenoidea: Polyonchoinea) and a gastrocotylinean pre-adult (Heteronchoinea) from pomacentrid and caesionid (Perciformes) fishes from Lizard Island, Great Barrier Reef, Australia
  50. Erratum to: 2017, 62(1) should be 2017, 62(2)
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