Anisakis pegreffii: A quantitative fluorescence PCR assay for detection in situ

• DOI: 10.1016/j.exppara.2010.11.008
• Publication Date: 2010-11-25T09:21:50Z
• Authors: Wenzhen Fang; Fan Liu; Shaolei Zhang; Junhua Lin; Shisan Xu; Damin Luo
• Keywords: Second internal transcribed spacer (ITS-2); 0301 basic medicine; 0303 health sciences; Fishes; Reproducibility of Results; Anisakiasis; TaqMan real-time PCR; DNA, Ribosomal; Polymerase Chain Reaction; Sensitivity and Specificity; Anisakis; In situ; Food-borne parasitic zoonosis; Fish Diseases; 03 medical and health sciences; Anisakis pegreffii; DNA, Ribosomal Spacer; Animals; 14. Life underwater; DNA Probes; Nematode; DNA Primers; Fluorescent Dyes

To facilitate improved diagnosis and detection of the third stage larva (L3) of Anisakis pegreffii from the Minnan-Taiwan bank fishing ground in Taiwan Strait, a real-time PCR method for the detection in situ and differentiation was developed to amplify a region of the second internal transcribed spacer (ITS-2) of this parasite. The real-time PCR assay was capable of detecting 1/3 of a single L3 in 30 mg of marine fish tissue, and also exhibited a high level of specificity for A. pegreffii, no fluorescence signals were observed in other five major larval anisakid species found in commercial marine fishes caught in this fishing ground.