Multiple primer PCR for the identification of anisakid nematodes from Taiwan Strait

• Published in: Experimental parasitology 2010-02, Vol.124 (2), p.197-201
• Main Authors: Fang, Wenzhen, Xu, Shisan, Zhang, Shaolei, Wang, Yinan, Chen, Xiaobin, Luo, Damin
• Format: Article
• Language: English
• Subjects: Animals; Anisakid nematodes; Anisakis; Anisakis physeteris; Ascaridida Infections - parasitology; Ascaridida Infections - veterinary; Ascaridoidea - classification; Ascaridoidea - genetics; Ascaridoidea - isolation & purification; Biological and medical sciences; Contracaecum; Contracaecum aduncum; DNA Primers - genetics; DNA, Helminth - chemistry; DNA, Helminth - isolation & purification; DNA, Ribosomal - chemistry; DNA, Ribosomal - isolation & purification; Fish Diseases - parasitology; Fishes; Fundamental and applied biological sciences. Psychology; Invertebrates; Life cycle. Host-agent relationship. Pathogenesis; Multiplex PCR; Nemathelminthia. Plathelmintha; Nematoda; Nuclear ribosomal DNA; PCR-RFLP; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; Protozoa; Raphidascaris; Seawater; Sequence Alignment; Species Specificity; Taiwan
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2009.09.011

There were six major larval anisakid species found in commercial marine fishes caught in the Minnan fishing ground in the Taiwan Strait: Anisakis physeteris, Anisakis pegreffii, Raphidascaris trichiuri, Contracaecum aduncum, Contracaecum muraenesoxi, Contracaecum sp. For rapid identification of the parasite species above, a single and a multiple primer PCR (multiplex PCR) method, using specific primers based on aligned sequences of the internal transcribed spacer ITS-1, 5.8S, and ITS-2 of nuclear ribosomal DNA, were jointly used for the rapid identification of these anisakid larvae. The primers yielded distinct PCR products for each of the anisakid nematodes, providing rapid and accurate tools for identifying anisakid nematodes with distinct geographical distribution.