Title: SPECIFICITY AND SENSITIVITY OF RANDOM AMPLIFIED POLYMORPHIC DNA ANALYSIS FOR THE IDENTIFICATION OF SINGLE LARVAE OF TRICHINELLA
Pozio, E - ROME, ITALY
Kapel, C - ROYAL VET & AG U, DENMARK
Submitted to: Journal of Parasitology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 14, 1998
Publication Date: N/A
Interpretive Summary: Trichinella is a nematode parasite which infects virtually all warm blooded animals. It is best known for causing disease in humans as a result of eating raw or undercooked meat, particularly pork. Control of Trichinella involves understanding the epidemiology of the parasite, especially sources of infection for pigs. The test described here provides a simple and rapid dmethod for determining the type of Trichinella found in pigs. Typing is part of the information needed to assess the source of infection (wild animals, rodents, other pigs). The test, which can be used on single worms will be very useful to veterinarians and public health workers who study the transmission and control of this parasite.
Single muscle larvae (ML) of Trichinella belonging to five species and two genotypes collected from 132 experimentally infected pigs and preserved in 75% ethyl alcohol at room temperature were identified by random amplified polymorphic DNA (RAPD) analysis to evaluate the specificity and sensitivity of this assay. A double blind protocol was used to assure that the interpretation of electrophoretic patterns after RAPD amplification was no influenced by an expected result. RAPD analysis of one larva from each sample resulted in an 88% match with the species or genotype that had been used to infect pigs, whereas the identification reached 100% specificity when an additional 1-4 ML were examined. Electrophoretic patterns obtained from larvae with partially damaged DNA (observed in 7 samples) complicated the identification, particularly for species and genotypes that are phylogenetically close each to other. The sensitivity of RAPD analysis is so great that the amount of DNA recovered from one muscle larva can be use for at least five assays.