Dideoxy fingerprinting: application to the genotyping of Echinococcus.
Dideoxy fingerprinting is an efficient method for the detection of sequence variation in PCR-amplified DNA segments. It is a hybrid between single-strand conformation polymorphism and dideoxy sequencing, employing only one dideoxynucleotide in the sequencing reaction. Herein, we report the application of dideoxy fingerprinting to genetically type cestodes of the genus Echinococcus, utilising the mitochondrial cytochrome c oxidase subunit I as the gene sequence for analysis. All of the seven genotypes (G1, G4, G6, G8, O, V and M2) examined could be readily differentiated from one another by their characteristic and reproducible dideoxy fingerprinting profiles. Only subtle variation in profiles was detected among some of the eight isolates representing genotype G1, and no variation was detected between two samples of genotype G4 and of genotype M2. The capacity of dideoxy fingerprinting to detect all nucleotide variations over 150-250bp fragments indicates that it should be possible to distinguish among all of the genotypes of Echinococcus thus far described. Although employed herein to display sequence variation in the cytochrome c oxidase subunit I of Echinococcus, dideoxy fingerprinting could be used for the high-resolution analysis of nucleotide variations in other parasite genes, without the need for DNA sequencing. This has important implications for studying the genetic structure of parasite populations.
Gasser
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RB
Department of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia. gasser@ariel.ucs.unimelb.edu.au
Zhu
X
X
McManus
D P
DP
D000818:Animals; D002162:Camelus / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D003670:Deer / Q000469:parasitology; D004285:Dogs / Q000469:parasitology; D004446:Echinococcus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000235:genetics*; D005838:Genotype; D006736:Horses / Q000469:parasitology; D006801:Humans; D007614:Macropodidae / Q000469:parasitology; D008928:Mitochondria / Q000201:enzymology; D016133:Polymerase Chain Reaction; D012377:Rodentia / Q000469:parasitology; D012756:Sheep / Q000469:parasitology
D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV