The random amplification of polymorphic DNA can discriminate species and strains of Echinococcus.
The random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), also called arbitrary priming PCR (AP-PCR), is a DNA polymorphism assay based on the amplification of genomic DNA using a single oligonucleotide primer of arbitrary nucleotide sequence. Using carefully controlled conditions, the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) proved useful for distinguishing Echinococcus species and genetically distinct strains of Echinococcus granulosus. Amongst twenty random 10 mer oligonucleotide primers tested, 4 (5'-GGGAATTCGG-3', 5'-GGCTGCAGAA-3', 5'-GGAGTACTGG-3', 5'-CCTCTAGACC-3') were most suitable for producing complex but characteristic patterns for discrimination purposes. Although the method amplifies vertebrate DNA, the resulting patterns do not interfere with the identification of Echinococcus isolates. Consequently, the technique can still be used effectively to distinguish Echinococcus species and strains even if host contamination occurs during the course of parasite collection. The approach complements several other methods, based on direct DNA examination, recently developed in our laboratory, which also allow unambiguous discrimination of Echinococcus genotypes.
Scott
J C
JC
Molecular Parasitology Unit, Queensland Institute of Medical Research, Brisbane, Australia.
McManus
D P
DP
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004446:Echinococcus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D005784:Gene Amplification; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D013045:Species Specificity
D017931:DNA Primers; D004247:DNA