Further molecular discrimination of Spanish strains of Echinococcus granulosus.
We have designed two polymerase chain reaction (PCR) primer sets (PEg9F1-PEg9R1 and PEg16F1-PEg16R1) and two PCR protocols (Eg9-PCR and Eg16-PCR) for discrimination of Echinococcus granulosus genotypes. The oligonucleotide sequences originate from two E. granulosus DNA multiplex-PCR amplification fragments, previously reported, that allows species-specific discrimination between Taenia saginata, Taenia solium, and E. granulosus. The Eg9-PCR, Eg16-PCR, and Eg9-PCR linked restriction fragment length polymorphism (RFLP) analysis was used to characterize 53 E. granulosus isolates from the central region of Spain, highly endemic for echinococcosis. The analysis resulted in: (i) the discrimination of E. granulosus from Echinococcus multilocularis; (ii) the characterisation and discrimination of discrete E. granulosus strains from Spain; and (iii) the identification of two distinct genotypes within E. granulosus Spanish pig isolates. To further characterize the genetic variants in pigs, fragments of the NADH dehydrogenase I (ND1) and the cytochrome c oxidase subunit I (CO1) genes were amplified from parasite DNA and sequenced. The results again revealed the presence of two distinct genotypes: the G1 (sheep-dog strain) and G7 (pig-dog strain) genotypes. This observation could have important consequences for human health in Spain. Furthermore, the Eg9-PCR, Eg16-PCR, and Eg9-PCR-RFLP protocols can be used as additional methods to discriminate various E. granulosus genotypes.
González
L M
LM
Instituto de Salud Carlos III, Centro Nacional de Microbiología, 28220 Majadahonda, Madrid, Spain. tgarate@isciii.es
Daniel-Mwambete
K
K
Montero
E
E
Rosenzvit
M C
MC
McManus
D P
DP
Gárate
T
T
Cuesta-Bandera
C
C
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000737:chemistry*; D004272:DNA, Mitochondrial / Q000737:chemistry; D004443:Echinococcosis / Q000469:parasitology*; D004446:Echinococcus / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000235:genetics; D005838:Genotype; D006736:Horses; D006801:Humans; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D012377:Rodentia; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012756:Sheep; D013030:Spain; D013045:Species Specificity; D013552:Swine
D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV