title: First molecular identification of the zoonotic parasite Anisakis pegreffii (Nematoda: Anisakidae) in a paraffin-embedded granuloma taken from a case of human intestinal anisakiasis in Italy.
abstract: BACKGROUND
  Anisakiasis is an important fish-borne zoonosis provoked by larval stages of nematodes belonging to the genus Anisakis. The detection and identification of human infections is difficult. This is due to: a) the low specificity of the clinical features and symptomatology related to human infections; b) the paucity of diagnostic features of larvae found in granulomatous lesions characteristic of "invasive anisakiasis"; and c) the lack morphological characters diagnostic at the specific level when larvae of Anisakis are detected. Thus, molecular-based diagnostic approaches are warranted.
  
  
  METHODS
  We have developed a PCR method that amplifies the DNA of Anisakis spp. in fixed paraffin-embedded tissues. This method was applied to a granuloma removed from a human case of intestinal anisakiasis in Italy. Specific primers of the mtDNA cox2 gene were used and sequence analysis was performed according to the procedures already established for species of Anisakis.
  
  
  RESULTS
  The sequence obtained (629 bp) was compared with those of the other species of Anisakis which have so far been genetically characterized and with sequences obtained from larval stages of Anisakis collected from the Mediterranean fish Engraulis encrasicolus. This enabled the genetic identification of the larva in the human tissue as A. pegreffii. This is the first instance of human intestinal anisakiasis diagnosed using PCR of DNA purified from a fixed eosinophilic granuloma embedded in paraffin.
  
  
  CONCLUSIONS
  The case of human anisakiasis presented reinforces the pathological significance of the species A. pegreffii to humans. The molecular/genetic methodological approach based on mtDNA cox2 sequence analysis, described here, can allow easy and rapid identification of Anisakis spp. in formalin-fixed and paraffin embedded tissues removed from cases of either gastric or intestinal human anisakiasis.
authors: Department of Public Health and Infectious Diseases, Sapienza - University of Rome, Ple Aldo Moro 5, 00185 Rome, Italy. simonetta.mattiucci@uniroma1.it
authors: Simonetta
authors: S
authors: Mattiucci
authors: Michela
authors: M
authors: Paoletti
authors: Francesco
authors: F
authors: Borrini
authors: Massimo
authors: M
authors: Palumbo
authors: Raffaele Macarone
authors: RM
authors: Palmieri
authors: Vincenzo
authors: V
authors: Gomes
authors: Alessandra
authors: A
authors: Casati
authors: Giuseppe
authors: G
authors: Nascetti
mesh_terms: D000818:Animals; D017129:Anisakiasis / Q000175:diagnosis / Q000469:parasitology*; D017163:Anisakis / Q000235:genetics* / Q000302:isolation & purification; D018509:DNA, Helminth / Q000235:genetics; D006099:Granuloma / Q000469:parasitology*; D006801:Humans; D007422:Intestines / Q000469:parasitology*; D007558:Italy; D008297:Male; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018509:DNA, Helminth