title: Development and validation of an inexpensive and efficient method for the extraction of Theileria orientalis DNA from blood.
abstract: Theileria orientalis is an emerging bovine pathogen in Australasia. PCR-based detection methods for this parasite are sensitive but relatively expensive, partly due to costs associated with DNA extraction. An inexpensive and efficient technique was developed for the extraction of T. orientalis DNA from blood based on hypotonic erythrocyte lysis and detergent-proteinase K treatment (DPK method). The DPK method compares favourably to a commercial extraction kit when paired with a T. orientalis multiplex qPCR.
authors: The ithree institute, University of Technology Sydney, Ultimo, NSW 2007, Australia; Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia.
authors: D R
authors: DR
authors: Bogema
authors: Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia.
authors: S A
authors: SA
authors: Fell
authors: Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia.
authors: B A
authors: BA
authors: O'Rourke
authors: Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia.
authors: D
authors: D
authors: Collins
authors: Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia.
authors: G J
authors: GJ
authors: Eamens
authors: Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries, Menangle, NSW 2568, Australia. Electronic address: cheryl.jenkins@dpi.nsw.gov.au.
authors: C
authors: C
authors: Jenkins
mesh_terms: D000818:Animals; D002417:Cattle; D016054:DNA, Protozoan / Q000097:blood* / Q000235:genetics; D003902:Detergents; D019286:Endopeptidase K; D004912:Erythrocytes; D015203:Reproducibility of Results; D016794:Theileria / Q000235:genetics*; D013801:Theileriasis / Q000097:blood / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D003902:Detergents; D019286:Endopeptidase K
keywords: DNA extraction; DPK method; Genotypes; Theileria orientalis; qPCR