Random amplified polymorphic DNA technique for the identification of Trichinella species.
The random amplified polymorphic DNA (RAPD) technique was successfully used to produce genetic fingerprints distinguishing between Trichinella spiralis and Trichinella britovi. The same patterns were obtained from purified and crude DNA preparations of pooled and single muscle larvae. RAPD fingerprinting was applied to muscle larvae preserved under different conditions and recovered from different hosts. Larvae recovered from fresh and frozen meat and stored at -20 degrees C for a long time or under 70% ethyl alcohol at room temperature for 30 d gave good and reproducible results. Single larvae recovered from a naturally infected wild boar and from a human biopsy gave fingerprints congruent to those obtained from T. britovi reference strains. The results prove that RAPD analysis is a quick method to distinguish between the autochthonous Trichinella species of Central-Southern Europe in less than 1 d after the detection of the infection. If necessary, the biological material can be frozen or stored under 70% ethyl alcohol at room temperature and sent to laboratories able to perform the RAPD analysis. The RAPD technique requires no prior knowledge of the molecular biology of the organism to be investigated and therefore appears to be a promising tool in parasitology for the identification of sibling species.
Bandi
C
C
Istituto di Patologia Generale Veterinaria, Università di Milano, Italy.
La Rosa
G
G
Comincini
S
S
Damiani
G
G
Pozio
E
E
D000818:Animals; D001483:Base Sequence; D016172:DNA Fingerprinting / Q000379:methods*; D006801:Humans; D007814:Larva / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles / Q000469:parasitology; D021141:Nucleic Acid Amplification Techniques; D011110:Polymorphism, Genetic; D013552:Swine; D014234:Trichinella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D017160:Trichinella spiralis / Q000235:genetics / Q000302:isolation & purification; D014235:Trichinellosis / Q000469:parasitology*