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Parasitology Research

, Volume 104, Issue 1, pp 79–84 | Cite as

Specific PCR assays for the identification of common anisakid nematodes with zoonotic potential

  • Q. Chen
  • H. Q. Yu
  • Z. R. Lun
  • X. G. Chen
  • H. Q. Song
  • R. Q. Lin
  • X. Q. Zhu
Original Paper

Abstract

Based on the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) for six taxa of anisakids, namely, Anisakis simplex (s.s.), Anisakis typica, Anisakis pegreffii, Hysterothylacium aduncum, Hysterothylacium sp, and Contracaccum osculatum C, specific primers were designed in the ITS-1 and/or ITS-2 for each of the six anisakid taxa. These specific primers were used to develop polymerase chain reaction (PCR) tools for the identification of these anisakid taxa of sea fish by amplifying partial ITS-1 and/or ITS-2 of rDNA from anisakid nematodes. This approach allowed their specific identification, with no amplicons being amplified from heterogeneous DNA samples, and sequencing confirmed the identity of the DNA fragments amplified. The minimum amounts of DNA detectable using the PCR assays were 0.5–1 ng. These PCR tools were then applied to ascertain the specific identity of 143 anisakid larval samples collected from fish in China, Canada, Thailand, and Indonesia, and these anisakid samples were identified to represent one of the six anisakid taxa. These PCR assays based on ITS sequences should provide useful molecular tools for the accurate identification and molecular epidemiological investigations of anisakid infections in humans and fish.

Keywords

Polymerase Chain Reaction Assay Specific Polymerase Chain Reaction Anisakiasis Anisakid Nematode Specific Polymerase Chain Reaction Product 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

Project support was provided by a grant from the Program for Changjiang Scholars and Innovative Research Team in University (Garnt No. IRT0723) to XQZ. The authors are grateful to Dr. M. Podolska of Sea Fisheries Institute, Poland for providing some anisakid samples from Poland. The experiments comply with the current laws of the countries in which the experiments were performed.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Q. Chen
    • 1
  • H. Q. Yu
    • 2
  • Z. R. Lun
    • 3
  • X. G. Chen
    • 4
  • H. Q. Song
    • 1
  • R. Q. Lin
    • 1
  • X. Q. Zhu
    • 1
  1. 1.College of Veterinary MedicineSouth China Agricultural UniversityGuangzhouPeople’s Republic of China
  2. 2.Laboratory of Animal QuarantineGuangzhou Entry–Exit Inspection and QuarantineZhujiang New City, GuangzhouPeople’s Republic of China
  3. 3.Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life SciencesSun Yat-Sen UniversityGuangzhouPeople’s Republic of China
  4. 4.Department of ParasitologySchool of Public Health and Tropical Medicine, Southern Medical UniversityGuangzhouPeople’s Republic of China

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