Molecular characterization of Trichinella genotypes by inter-simple sequence repeat polymerase chain reaction (ISSR-PCR).
A bulk analysis of inter-simple sequence repeat-polymerase chain reaction (ISSR-PCR) provides a quick, reliable, and highly informative system for DNA banding patterns that permit species identification. The present study evaluates the applicability of this system to Trichinella species identification. After a single amplification carried out on a single larva with the primer 816([CA]nRY) under high stringency conditions, which provide high reproducibility, we were able to identify by consistent banding patterns 5 sibling species: Trichinella spiralis (ISS48), 2 Trichinella britovi isolates (ISS11 and ISS86), Trichinella murrelli (ISS35), Trichinella nativa (ISS71), Trichinella nelsoni (ISS29); 3 additional Trichinella genotypes: T8 (ISS149), T9 (ISS408 and ISS409), and T6 (ISS34); and the nonencapsulated species Trichinella pseudospiralis (ISS13). Moreover, 33 new Trichinella isolates from 2 zoogeographical regions were unequivocally identified. All Trichinella isolates have shown an identical pattern with those produced by the reference strain. According to these data, we have demonstrated that ISSR-PCR is a robust technique that emerges as a useful new application for the molecular identification of Trichinella isolates in epidemiological studies.
Fonseca-Salamanca
F
F
Unidad de Parasitología, Facultad de Medicina, Universidad de La Frontera, Casilla, Temuco, Chile.
Nogal-Ruiz
J J
JJ
Benito
C
C
Camachot
M V
MV
Martínez-Fernández
A R
AR
D000818:Animals; D046028:Canidae; D017931:DNA Primers / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005260:Female; D005838:Genotype; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D034421:Sus scrofa; D014234:Trichinella / Q000145:classification / Q000235:genetics*
D017931:DNA Primers; D018509:DNA, Helminth