title: Molecular discrimination of Echinococcus granulosus and Echinococcus multilocularis by sequencing and a new PCR-RFLP method with the potential use for other Echinococcus species.
abstract: OBJECTIVE
  To develop a novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol using a new genomic marker sequence and a novel set of restriction enzymes in order to detect and discriminate 2 Echinococcus species, E. granulosus and E. multilocularis, found in formalin-fixed paraffin-embedded (FFPE) human tissues.
  
  
  METHODS
  DNA was isolated from 11 FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis. A mitochondrial genomic marker region was amplified and sequenced using a novel primer pair and a new PCR-RFLP protocol was developed for the detection and discrimination of E. granulosus and E. multilocularis using a set of restriction enzymes including AccI, MboI, MboII, and TsoI.
  
  
  RESULTS
  The selected marker region was amplified using DNA isolated from FFPE human tissue samples positive for cystic echinococcosis or alveolar echinococcosis and the discrimination of E. granulosus and E. multilocularis was accomplished by use of the novel PCR-RFLP method.
  
  
  CONCLUSIONS
  In this PCR-RFLP protocol, use of any single restriction enzyme is enough for the discrimination of E. granulosus and E. multilocularis. The PCR-RFLP protocol can be potentially used for the discrimination of 5 other Echinococcus species: E. oligarthus, E. shiquicus, E. ortleppi, E. canadensis, and E. vogeli.
authors.forename: Çağrı
authors.initials: Ç
authors.lastname: Şakalar
authors.forename: Salih
authors.initials: S
authors.lastname: Kuk
authors.forename: Ahmet
authors.initials: A
authors.lastname: Erensoy
authors.forename: Adile Ferda
authors.initials: AF
authors.lastname: Dağli
authors.forename: İbrahim Hanifi
authors.initials: İH
authors.lastname: Özercan
authors.forename: Ülfet
authors.initials: Ü
authors.lastname: Çetınkaya
authors.forename: Süleyman
authors.initials: S
authors.lastname: Yazar
mesh_terms: D000818:Animals; D018509:DNA, Helminth / Q000302:isolation & purification; D004272:DNA, Mitochondrial / Q000302:isolation & purification; D004443:Echinococcosis / Q000235:genetics* / Q000469:parasitology; D004444:Echinococcosis, Hepatic / Q000235:genetics / Q000469:parasitology; D048209:Echinococcus granulosus / Q000235:genetics / Q000302:isolation & purification*; D048210:Echinococcus multilocularis / Q000235:genetics / Q000302:isolation & purification*; D005557:Formaldehyde; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D005557:Formaldehyde