[Application of the PCR-RFLP technique for the species-specific identification of nematodes involved in human anisakiasis].
OBJECTIVE
We intended to develop a molecular test allowing a species-specific identification of the anisakid human parasite independently of its evolutive stage.


METHODS
Anisakid larvae were obtained from fish destined to human consumption. In the PCR-RFLP test, the DNA corresponding to the ITS-1, 5.8S rRNA gene, ITS-2 and approximately 70 of the 28S rRNA gene region was amplified and a positive control was included. Products were digested with the endonuclease Taq I and subsequently sequenced.


RESULTS
Different anisakids (Anisakis simplex/Hysterothylacium aduncum) yield different amplification (960 and 1,010 bp fragments) and restriction patterns, which were in accord with previously described patterns of the same parasites from others geographical regions.


CONCLUSIONS
The high sensitivity of the test and the absence of intraspecies variations confirms the utility of this assay in the identification of human parasites involved in human anisakiasis including larvae from resected biopsies.
Perteguer
María Jesús
MJ
Servicio de Parasitología, Instituto de Salud Carlos III, Centro Nacional de Microbiología, Ctra. Majadahonda-Pozuelo, km 2,2.28220 Majadahonda, Madrid, Spain.
Ortiz
Guadalupe
G
García
Elena
E
Flores
María
M
Rodríguez
Esperanza
E
Ubeira
Florencio M
FM
Gárate
Teresa
T
D000818:Animals; D017129:Anisakiasis / Q000175:diagnosis; D017163:Anisakis / Q000145:classification* / Q000302:isolation & purification; D018509:DNA, Helminth / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005399:Fishes / Q000469:parasitology*; D005506:Food Contamination; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
D018509:DNA, Helminth