Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지

Differential diagnosis of Trichostrongylus and hookworm eggs via PCR using ITS-1 sequence

  • Yong, Tai-Soon (Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine) ;
  • Lee, Jong-Ho (Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine) ;
  • Sim Seo-Bo (Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine) ;
  • Lee, Jong-Weon (Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine) ;
  • Min Duk-Young (Department of Parasitology, Hanyang University College of Medicine) ;
  • Chai, Jong-Yil (Department of Parasitology, Seoul National University College of Medicine, and Institute of Endemic Diseases, Seoul National University Medical Research Center) ;
  • S. Eom, Kee-Seon (Department of Parasitology and Medical Research Institute, Chungbuk National University College of Medicine) ;
  • Sohn Woon-Mok (Department of Parasitology and Institute of Health Sciences, Gyeongsang National University College of Medicine) ;
  • Lee, Soon-Hyung (Department of Parasitology, Seoul National University College of Medicine, and Institute of Endemic Diseases, Seoul National University Medical Research Center) ;
  • Rim, Han-Jong (Department of Parasitology, Korea University College of Medicine)
  • Published : 2007.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Trichostrongylus eggs observed in cellophane-thick smears are difficult, in practice, to distinguish from hookworm eggs. In order to overcome these limitations, a molecular approach was conducted. A Trichostrongylus colubriformis adult worm was obtained from a human in Laos, which was identified morphologically. ITS-1 sequence of this worm was determined, and found to be most similar with that of T. colubriformis among the Trichostrongylus spp. reported so far. Then, this sequence was compared with those of human hookworm species, Ancylostoma duodenale and Necator americanus, and species-specific oligonucleotide primers were designed. Polymerase chain reaction(PCR) using these primers evidenced specifically amplified PCR products of Trichostrongylus sp., A. duodenale and N. americanus from the eggs of each(520 bp, 690 bp, and 870 bp, respectively). A species-specific PCR technique can be developed in order to study the epidemiology of Trichostrongylus spp. and hookworms in endemic areas.

Keywords

References

  1. Beaver PC, Jung RC, Cupp EW (1984) Clinical Parasitology. 9th ed. pp 289-291, Lea & Febiger, Philadelphia, USA
  2. Chilton NB, Gasser RB (1999) Sequence differences in the internal transcribed spacers of DNA among four species of hookworm (Ancylostomatoidea: Ancylostoma). Int J Parasitol 29: 1971-1977 https://doi.org/10.1016/S0020-7519(99)00148-4
  3. de Gruijter JM, van Lieshout L, Gasser RB, Verweij JJ, Brienen EA, Ziem JB, Yelifari L, Polderman AM (2005) Polymerase chain reaction-based differential diagnosis of Ancylostoma duodenale and Necator american us infections in humans in northern Ghana. Trop Med Int Health 10: 574-580 https://doi.org/10.1111/j.1365-3156.2005.01440.x
  4. Gasser RB, Stewart LE, Speare R (1996) Genetic markers in ribosomal DNA for hookworm identification. Acta Trop 62: 15-21 https://doi.org/10.1016/S0001-706X(96)00015-0
  5. Ghadirian E, Arfaa F (1975) Present status of trichostrongyliasis in Iran. Am J Trop Med Hyg 24: 935-941 https://doi.org/10.4269/ajtmh.1975.24.935
  6. John DT, Petri WA (2006) Markell and Voge's Medical Parasitology. 9th ed. pp 266-267, Elsevier, St. Louis, USA
  7. Monti JR, Chilton NB, Qian BZ, Gasser RB (1998) Specific amplification of Necator americanus or Ancylostoma duodenale DNA by PCR using markers in ITS-1 rDNA, and its implications. Mol Cell Probes 12: 71-78 https://doi.org/10.1006/mcpr.1997.0151
  8. Rim HJ, Chai JY, Min DY, Cho SY, Eom KS, Hong SJ, Sohn WM, Yong TS, Deodato G, Standgaard H, Phommasack B, Yun CH, Hoang EH (2003) Prevalence of intestinal parasite infections on a national scale among primary schoolchildren in Laos. Parasitol Res 91: 267-272 https://doi.org/10.1007/s00436-003-0963-x
  9. Romstad A, Gasser RB, Nansen P, Polderman AM, Chilton NB (1998) Necator americanus (Nematoda: Ancylostomatidae) from Africa and Malaysia have different ITS-2 rDNA sequences. Int J Parasitol 28: 611-615 https://doi.org/10.1016/S0020-7519(97)00213-0
  10. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_Xwindows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 4876-4882 https://doi.org/10.1093/nar/25.24.4876
  11. Verweij JJ, Polderman AM, Wimmenhove MC, Gasser RB (2000) PCR assay for the specific amplification of Oesophagostomum bifurcum DNA from human faeces. Int J Parasitol 30: 137-142 https://doi.org/10.1016/S0020-7519(99)00175-7
  12. Zhan B, Li T, Xiao S, Zheng F, Hawdon JM (2001) Speciesspecific identification of human hookworms by PCR of the mitochondrial cytochrome oxidase I gene. J Parasitol 87: 1227-1229 https://doi.org/10.1645/0022-3395(2001)087[1227:SSIOHH]2.0.CO;2